From owner-repertoires@net.bio.net Wed Jul 02 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Todd L Graybill Newsgroups: bionet.molecules.repertoires Subject: Bead Integrity Date: 3 Jul 1997 10:05:25 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 33 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <9707031921.AB6526@pho903.sbphrd.com> Reply-To: Todd L Graybill NNTP-Posting-Host: net.bio.net [I have reposted this message from MOL-DIVERSITY as it appears to be interesting for molreps readers also - AW] In one forum or another, the frailty of resin beads has been a topic of discussion. For example, it has become quite well known that stirring suspensions of resins with magnetic stirbars typically leads to loss of bead integrity as a result of bead fragmentation. Fragmentation can result in frit clogging or loss of resin from teabags or IRORI microKans. It is also important to avoid bead fragmentation if manipulation of single beads is required for library analysis or screening. I would like to initiate a discussion concerning bead fragmentation among this MOL-DIVERSITY Group. Have you encountered bead fragmentation? Was it a mechanical process such as magnetic stirring that caused it or was it the result of a non-mechanical process such as a chemical reaction, shrink/swell resin washing etc.? In particular, has anyone reported data to document whether rapid changes in solvent polarity (MeOH/DCM or THF/water) during shrink/swell resin washings cause fragmentation or loss of bead integrity? Are larger beads more prone to fragmentation than smaller beads? Is one polymer bead (i.e. polystyrene vs PEG-based) more prone than another? Other comments or observations are welcomed and encouraged. Todd L. Graybill SmithKline Beecham Pharmaceuticals Medicinal Chemistry Tel. 610-270-6333 Email Todd_L_Graybill@sbphrd.com From owner-repertoires@net.bio.net Thu Jul 03 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Vic Ilag Newsgroups: bionet.molecules.repertoires Subject: M13 anti-pVIII Ab Date: 4 Jul 1997 06:31:42 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 9 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <5pioi5$o9n@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net Does anyone know of a vendor selling a monoclonal/polyclonal to the pVIII protein of filamentous phage (M13, fd, f1, etc) or labs that are willing to provide it. Thanks in advance. Please let me know. vic From owner-repertoires@net.bio.net Tue Jul 08 23:00:00 1997 Path: biosci!biosci!not-for-mail From: M.Foley@latrobe.edu.au (Dr Michael Foley) Newsgroups: bionet.molecules.repertoires Subject: searching databases Date: 9 Jul 1997 01:59:43 -0700 Organization: La Trobe University Lines: 14 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <5pv5k7$ont$1@news.latrobe.edu.au> NNTP-Posting-Host: net.bio.net Hi All, If after panning some random peptide libraries and you want to search the sequence in the databases, does anyone have a feeling for what the best approach is? We search using blast via the web, but I was wondering what sort of parameters you might require to get realistic homologies with such short sequences. Is anyone doing this and would like to comment. Cheers, Mick From owner-repertoires@net.bio.net Tue Jul 08 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Andrew Wallace Newsgroups: bionet.molecules.repertoires Subject: UK Postdoc Cancer Research Date: 9 Jul 1997 08:03:06 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 33 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <5q07us$575@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net Research Fellow School of Biology and Biochemistry This post, funded by the Association for International Cancer Research, is available from 1 October for three years, to assist in a research project carrying out work on the 67 kDa laminin receptor. The project will use phage display techniques to disclose novel receptor ligands and will have applications in angiogenesis and cancer biology. Applicants must have a minimum 2.2 honours degree or equivalent, preferably 2.1 or better or equivalent, in a scientific subject and a PhD in molecular biology or biochemistry as well as experience of recombinant DNA methods including cloning and expression of DNA fragments in bacterial vectors. The following is desirable: experience in working with phage display libraries and mammalian cell culture together with a good publication record. Salary range: 16,927 - 17,606 pounds Sterling per annum. Applicants, quoting Ref: 97/F154C, may obtain further particulars from the Personnel Office, The Queen's University of Belfast, BT7 1NN, Tel (+441232) 273246/273044 or FAX (+441232) 324944. Closing date: 22 August 1997. ------------------------------------------------------------------ Andrew Wallace, Ph.D, Queens University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html From owner-repertoires@net.bio.net Tue Jul 08 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Andrew Wallace Newsgroups: bionet.molecules.repertoires Subject: Updated list of meetings and courses Date: 9 Jul 1997 07:20:03 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 152 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <5q065u$2uf@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net Meetings and courses on combinatorial libraries and phage display. ***************************************************************** _________________________________________________________________ *** MEETINGS *** _________________________________________________________________ Combinatorial Approaches to Chemistry and Biology 27-30 July 1997 Churchill College, Cambridge, UK Organised by the Royal Society of Chemistry Closing date for applications 13 June 1997 Contact: Elaine Wellingham Conference Secretariat Field End House Bude Close Nailsea Bristol BS19 2FQ UK Tel. & Fax: +44 (0)1275 853311 Email: confsec@dial.pipex.com _________________________________________________________________ IBC's 2nd International Exposition and Symposium Drug Discovery Technology `97 Integrating R&D Activities to Accelerate Drug Development August 11-14, 1997 * Sheraton San Diego Hotel and Marina * San Diego, CA For more information about IBC or to register for this conference, please contact IBC USA Conferences, 225 Turnpike Road, Southborough, MA 01772-1749, USA. Tel: (508)481-6400 Fax: (508)481-7911 E-mail: inq@ibcusa.com Internet: http://www.ibcusa.com/conf/drugdisc _________________________________________________________________ Cambridge Healthtech Institutes First European Conference on Combinatorial Chemistry and High Throughput Screening December 4-5, 1997 Hotel Arts Barcelona Barcelona, Spain Contact: Irene Phelan, Conference Director Phone: 617-630-1362 Fax: 617-630-1325 e-mail: iphelan@healthtech.com Organized by Cambridge Healthtech Institute 1037 Chestnut Steet, Newton Upper Falls, MA 02164 USA http://www/healthtech.com/conferences/ _________________________________________________________________ IBC's 2nd Annual Conference on Display Technologies: Advances in Protein Engineering, Drug/Vaccine Development and Molecular Evolution; August 4-5, 1997-Boston, MA For More Information: Call IBC USA Conferences at 508-481-6400, Fax: 508-481-4473 or email aheghinian@ibcusa.com for complete conference brochure or Register Online at http://www.ibcusa.com/conf/display _________________________________________________________________ IBC's 3rd Interactive "Nuts & Bolts" Forum Combinatorial Technologies Case Studies, Practical Experiences and Detailed Results October 23-24, 1997 * Hotel del Coronado * Coronado, CA Organized and Sponsored by: IBC USA Conferences 225 Turnpike Road Southborough, MA 02132 Ph: (508)481-6400 Fax: (508)481-7911 E-mail: inq@ibcusa.com http://www.ibcusa.com/conf/combinatorial _________________________________________________________________ Phage Display of Proteins and Peptides Biochemical Society Meeting University College Galway 3-5 September 1997 Deadline for poster registration 6 June 1997 Further information from: Michelle Mandale Meetings Office The Biochemical Society 59 Portland Place London W1N 3AJ Fax: (+44)171-637-7626 email: meetings@biochemsoc.org.uk __________________________________________________________________ The 8th Cyprus Conference on New Methods in Drug Research Limassol, Cyprus May 4-9, 1998 Conference Executive: Chair: Alex Makriyannis (U. Conn. Storrs, USA) Committee: N. Castagnoli (USA), J.P. Devlin (USA), U. Hacksell (Sweden), L.H. Hurley (USA) and M. Abou-Gharbia (USA) Conference Location Time and Registration: The Conference will take place from Monday, May 4th through Friday, May 9th 1998 at the five-star Hawaii Beach Hotel, Limassol, Cyprus. Attendance is restricted to 150 participants to encourage interaction. Registration fees ($595 per person; $395 for academics) includes attendance at all sessions and morning and afternoon breaks, but not meals. Sponsor: The European Federation of Medicinal Chemistry Contact: ARRT International, Inc., P.O. Box 1838, New Milford, CT 06776, USA Telephone: 860-355-5195; Facsimile: 860-355-5975; e-Mail: arrtintl@prodigy.net _________________________________________________________________ *** COURSES *** _________________________________________________________________ PHAGE DISPLAY OF COMBINATORIAL ANTIBODY LIBRARIES Cold Spring Harbor Laboratory November 4 -17 , 1997 APPLICATION DEADLINE: July 15, 1997 Organisers: Carlos Barbas, Scripps Research Institute Dennis Burton, Scripps Research Institute Gregg Silverman, University of California, San Diego For Additional Information, please contact: Cold Spring Harbor Laboratory The Meetings & Courses Office 1 Bungtown Road, PO Box 100 Cold Spring Harbor, NY 11724-2213 USA Phone: (+1)516-367-8346 Fax: (+1)516-367-8845 email: meetings@cshl.org __________________________________________________________________ ------------------------------------------------------------------ Andrew Wallace, Ph.D, Queens University Belfast, N. Ireland (UK) a.wallace@qub.ac.uk http://web.qub.ac.uk/bb/awpage/wallace.html From owner-repertoires@net.bio.net Tue Jul 08 23:00:00 1997 Path: biosci!biosci!not-for-mail From: "Dimitris K. Agrafiotis" Newsgroups: bionet.molecules.repertoires Subject: Review available Date: 9 Jul 1997 05:01:51 -0700 Organization: 3-Dimensional Pharmaceuticals, Inc. Lines: 13 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <33C2AB7D.3F18@3dp.com> Reply-To: "Dimitris K. Agrafiotis" NNTP-Posting-Host: net.bio.net I have just completed a ~40-page review on Molecular Diversity for the Encyclopedia of Computational Chemistry to be published later this year by Wiley. If you would like a preprint, please drop me a note. -- Dimitris K. Agrafiotis, PhD | E-mail: dimitris@3dp.com 3-Dimensional Pharmaceuticals, Inc. | Tel: (610) 458-6045 665 Stockton Drive, Suite 104 | Fax: (610) 458-8249 Exton, PA 19341 From owner-repertoires@net.bio.net Tue Jul 15 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Shu-Kun Lin Newsgroups: bionet.molecules.repertoires Subject: ECSOC-1, call for papers Date: 16 Jul 1997 02:17:00 -0700 Organization: University of Basel, Switzerland Lines: 41 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <01IL4JMH19228Y665C@ubaclu.unibas.ch> Reply-To: Shu-Kun Lin NNTP-Posting-Host: net.bio.net The First International Electronic Conference on Synthetic Organic Chemistry, ECSOC-1, September 1-30, 1997 (http://www.unibas.ch/mdpi/ecsoc/ in Europe and http://www.mdpi.org/ecsoc/ in the USA) ECSOC is diveided into several sections. I am responsible for its section F. Information and Compound Archives Management: Molecular diversity assessment, compound archives, databases. Contributions are now sought in the areas of Molecular diversity assessment, compound archives, databases management. Please submit your papers in the area of molecular diversity to me soon, as the deadline of July 15, 1997 is approaching. ECSOC-1 section B is also of great interest to us: B. Combinatorial Synthesis and Automation: New technologies for synthesis of oligomers and small molecules libraries. Please send your papers directly to the section convenor Dr. Eduard Felder, Pharmaceuticals Division, Novartis, 4002 Basel, Switzerland, phone +41 61 696 5293, fax +41 61 696 6174, e-mail Eduard.Felder@chbs.mhs.ciba.com Shu-Kun --------------------------------------------------------- Dr. Shu-Kun Lin Molecular Diversity Preservation International (MDPI) Saengergasse 25, CH-4054 Basel, Switzerland ECSOC-1: www.mdpi.org/ecsoc-1.htm e-mails: Lin@ubaclu.unibas.ch Lin@mdpi.org Tel. +41 79 322 3379, Fax +41 61 302 8918 -------------------------------------------------------- From owner-repertoires@net.bio.net Tue Jul 15 23:00:00 1997 Path: biosci!biosci!not-for-mail From: PETER WILLETT Newsgroups: bionet.molecules.repertoires Subject: Call for papers Date: 16 Jul 1997 02:14:23 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 35 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <2A4A1781824@ramsley.shef.ac.uk> Reply-To: PETER WILLETT NNTP-Posting-Host: net.bio.net CALL FOR PAPERS: COMPUTATIONAL APPROACHES TO THE DESIGN AND ANALYSIS OF COMBINATORIAL LIBRARIES The Molecular Graphics and Modelling Society and the Chemical Structure Association are organising a joint conference on the computational analysis of molecular diversity, to be held in Sheffield, UK 14-16 April 1998. Contributions are now sought in areas such as: the design of diverse and focussed libraries; algorithms and methods for the selection of database subsets; the representation and searching of combinatorial libraries and analogous virtual databases; the design and use of both in-house and commercial software for diversity analysis; diversity indices; compararison of property-based and fingerprint-based (dis)similarity measures; and the integration of diversity analysis with other computational tools, such as docking and QSAR. People wishing to present their work should submit a 500-word abstract (preferably by email) to Dr V.J. Gillet, Department of Information Studies, University of Sheffield, Sheffield S10 2TN, UK (tel. 044-114-2222652, fax 044-1142780300, email v.gillet@sheffield.ac.uk) by 1st October 1997. Professor Peter Willett, Department of Information Studies, University of Sheffield, Sheffield S10 2TN, U.K. Telephone: (0)114-2222633 (0)114-2222630 (for the general office) Fax: (0)114-2780300 Web: http://www.shef.ac.uk/uni/academic/I-M/is/lecturer/peter.html From owner-repertoires@net.bio.net Tue Jul 22 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Stephen Donoghue Newsgroups: bionet.molecules.repertoires Subject: Pharmacia phage display Date: 23 Jul 1997 00:54:56 -0700 Organization: Pathology Department , Leicester, U.K. Lines: 18 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <33D3868A.454F@le.ac.uk> NNTP-Posting-Host: net.bio.net Hello, I'm about to board the phage display train and i've bought the Pharmacia kit. I was wondering how effective other users find it. In addition, i was wondering if anybody knows the sequence of light and heavy primers which are initially used to amplify the respective variable genes. Mucho thanko in advance. Stephen Donoghue Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX email sed7@le.ac.uk From owner-repertoires@net.bio.net Tue Jul 22 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Michal Opas Newsgroups: bionet.molecules.repertoires Subject: Calreticulin Meeting: 1st circular [repost] Date: 23 Jul 1997 03:40:24 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 325 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Dear Colleague, We are delighted to announce that Calreticulin Workshop, devoted to the structure and function of calreticulin and related proteins, will take place on March 31 - April 2, 1998 in Banff, Alberta, Canada. The Workshop will provide unique opportunity to meet and interact with the scientists interested in calreticulin research in spectacular surroundings of Banff National Park in Canadian Rocky Mountains. We are sure that the Banff Calreticulin Workshop will be an important forum to share the latest findings and to develop future interactions. Calreticulin has been implicated to play a role in almost every aspect of cell biology as outlined in a brief overview below. We hope that the Workshop will be useful to sort out some of the latest discoveries and controversies concerning calreticulin and implication of this protein in a variety of biological systems. On the behalf of the Organizing Committee we would like to invite you to participate in the Workshop. The Calreticulin Workshop is a satellite meeting to the 8th Fisher Winternational Symposium on Cellular and Molecular Biology which will be held April 2-5, 1998, also at the Banff Conference Centre. The Winternational Symposium, which is co-sponsored by our Society and Fisher Scientific, is held annually, with a different focus each year. The theme for the 1998 meeting is : "Membrane Proteins in Health and Disease." Further information about the Fisher Winternational or the satellite meetings can be obtained by visiting the Society's web site at http://www.csbmcb.ca/english/bulletin/winternational.e.html or by contacting the Chair of the Scientific Program Committee by E-mail: Carol Cass . I hope you will participate in the Calreticulin Workshop. If you would like to receive further information please send a request (if possible, by e-mail) to Michal Opas at: m.opas@utoronto.ca or at: Department of Anatomy & Cell Biology University of Toronto Medical Sciences Building Toronto, Ontario, M5S 1A8 Canada tel: (416) 978-8947 fax: (416) 978-3954 I look forward to hearing from you in the near future. For The Organizing Committee Sincerely yours Michal Opas Calreticulin, a multifunctional protein Calreticulin, 60 kDa Ca-binding protein [1], is a major component of the endoplasmic reticulum (ER) of non-muscle cells [2-7]. The protein is of high physiological importance as it knockout is embryonic lethal [8]. Along with a wide tissue distribution [9], calreticulin is present in diverse animal and plant species [10]. calreticulin is a resident ER protein as demonstrated by a variety of biochemical and immunological techniques [1,3,4,6,11]. The protein is synthesized with an N-terminal signal sequence and it terminates with the KDEL sequence [3,12] which is responsible for retrieval of proteins to the lumen of the ER [13,14]. Calreticulin functions in vivo as a Ca storage protein [15,16]. It also has been well established that calreticulin is a chaperone [17-21] and it shows similarity in amino acid sequence to a part of calnexin, an ER membrane chaperone [22]. The Ca storage and chaperone functions of calreticulin are consistent with both the ER localization of calreticulin and its structure. Stable overexpression of calreticulin increases both cell-substratum and cell-cell adhesiveness with concomitant upregulation of adhesion-specific cytoskeletal protein, vinculin [23]. Upregulation of calreticulin also affects adhesion-dependent phenomena such as cell motility (which decreases) and cell spreading (which increases). Downregulation of calreticulin brings about inverse effects. In addition to the Ca storage and chaperone function, calreticulin modulates gene expression [24,25]. In vitro, calreticulin interaction with the DNA binding domain of the glucocorticoid receptor prevents the receptor from interacting with its glucocorticoid response element [24]. Transcriptional activation by glucocorticoid and androgen receptors in vivo is inhibited in cells overexpressing full length calreticulin [24,25]. Calreticulin itself is stress-regulated by heat and heavy metals [26-28]. Calreticulin has antithrombotic activity [29]. A host of other putative calreticulin functions includes a role in autoimmune diseases [30-34]. The protein affects replication of the Rubella virus RNA [35,36]. In cytolytic T lymphocytes it is found in the lytic granules where it may play a role in killing of target cells [37]. In human neutrophils calreticulin may contribute to the process of phagocytosis [38]. In line with the reported functional diversity, calreticulin was reported to be present in most cellular compartments [10,11,37,39,40], including the outer cell surface [41,42]. Recent hypotheses regarding calreticulin function have been presented by Krause and Michalak [43]. References 1. Ostwald TJ, MacLennan DH: Isolation of a high affinity calcium binding protein from sarcoplasmic reticulum. J Biol Chem 1974, 249:974-979. 2. Baksh S, Michalak M: Expression of calreticulin in Escherichia coli and identification of its Ca2+ binding domains. J Biol Chem 1991, 266:21458-21465. 3. Fliegel L, Burns K, Opas M, Michalak M: The high-affinity calcium binding protein of sarcoplasmic reticulum. Tissue distribution, and homology with calregulin. Biochim Biophys Acta 1989, 982:1-8. 4. Opas M, Dziak E, Fliegel L, Michalak M: Regulation of expression and intracellular distribution of calreticulin, a major calcium binding protein of nonmuscle cells. J Cell Physiol 1991, 149:160-171. 5. Milner RE, Baksh S, Shemanko C, Carpenter MR, Smillie L, Vance JE, Opas M, Michalak M: Calreticulin, and not calsequestrin, is the major calcium binding protein of smooth muscle sarcoplasmic reticulum and liver endoplasmic reticulum. J Biol Chem 1991, 266:7155-7165. 6. Michalak M, Baksh S, Opas M: Identification and immunolocalization of calreticulin in pancreatic cells: no evidence for "calciosomes". Exp Cell Res 1991, 197:91-99. 7. Michalak M, Milner RE, Burns K, Opas M: Calreticulin. Biochem J 1992, 285:681-692. 8. Coppolino MG, Woodside MJ, Demaurex N, Grinstein S, St-Arnaud R, Dedhar S: Calreticulin is essential for integrin-mediated calcium signalling and cell adhesion. Nature 1997, 386:843-847. 9. Tharin S, Dziak E, Michalak M, Opas M: Widespread tissue distribution of rabbit calreticulin, a non-muscle functional analogue of calsequestrin. Cell Tissue Res 1992, 269:29-37. 10. Opas M: The intracellular distribution and expression of calreticulin. In Calreticulin, edited by Michalak M. Georgetown: R.G. Landes; 1996:31-41. 11. Koch GLE: The endoplasmic reticulum and calcium storage. BioEssays 1990, 12:527-531. 12. Fliegel L, Burns K, MacLennan DH, Reithmeier RAF, Michalak M: Molecular cloning of the high affinity calcium-binding protein (calreticulin) of skeletal muscle sarcoplasmic reticulum. J Biol Chem 1989, 264:21522-21528. 13. Pelham HRB: Control of protein exit from the endoplasmic reticulum. Annu Rev Cell Biol 1989, 5:1-23. 14. S=F6nnichsen B, F=FCllekrug J, Van PN, Diekmann W, Robinson DG, Mieskes G: Retention and retrieval: Both mechanisms cooperate to maintain calreticulin in the endoplasmic reticulum. J Cell Sci 1994, 107:2705-2717. 15. Bastianutto C, Clementi E, Codazzi F, Podini P, De Giorgi F, Rizzuto R, Meldolesi J, Pozzan T: Overexpression of calreticulin increases the Ca2+ capacity of rapidly exchanging Ca2+ stores and reveals aspects of their lumenal microenvironment and function. J Cell Biol 1995, 130:847-855. 16. Liu N, Fine RE, Simons E, Johnson RJ: Decreasing calreticulin expression lowers the Ca2+ response to bradykinin and increases sensitivity to ionomycin in NG-108-15 cells. J Biol Chem 1994, 269:28635-28639. 17. Nauseef WM, McCormick SJ, Clark RA: Calreticulin functions as a molecular chaperone in the biosynthesis of myeloperoxidase. J Biol Chem 1995, 270:4741-4747. 18. Wada I, Imai S, Kai M, Sakane F, Kanoh H: Chaperone function of calreticulin when expressed in the endoplasmic reticulum as the membrane-anchored and soluble forms. J Biol Chem 1995, 270:20298-20304. 19. Nigam SK, Goldberg AL, Ho S, Rhode MF, Bush KT, Sherman MY: A set of endoplasmic reticulum proteins possessing properties of molecular chaperones includes Ca2+-binding proteins and members of the thioredoxin superfamily. J Biol Chem 1994, 269:1744-1749. 20. Otteken A, Moss B: Calreticulin interacts with newly synthesized human immunodeficiency virus type 1 envelope glycoprotein, suggesting a chaperone function similar to that of calnexin. J Biol Chem 1996, 271:97-103. 21. Hebert DN, Foellmer B, Helenius A: Calnexin and calreticulin promote folding, delay oligomerization and suppress degradation of influenza hemagglutinin in microsomes. EMBO J 1996, 15:2961-2968. 22. Bergeron JJM, Brenner MB, Thomas DY, Williams DB: Calnexin: a membrane-bound chaperone of the endoplasmic reticulum. Trends Biochem Sci 1994, 19:124-128. 23. Opas M, Szewczenko-Pawlikowski M, Jass GK, Mesaeli N, Michalak M: Calreticulin modulates cell adhesiveness via regulation of vinculin expression. J Cell Biol 1996, 135:1913-1923. 24. Burns K, Duggan B, Atkinson EA, Famulski KS, Nemer M, Bleackley RC, Michalak M: Modulation of gene expression by calreticulin binding to the glucocorticoid receptor. Nature 1994, 367:476-480. 25. Dedhar S, Rennie PS, Shago M, Leung-Hagesteijn C-Y, Yang H, Filmus J, Hawley RG, Bruchovsky N, Cheng H, Matusik RJ, Gigu=E8re V: Inhibition of nuclear hormone receptor activity by calreticulin. Nature 1994, 367:480-483. 26. Nguyen TQ, Capra JD, Sontheimer RD: Calreticulin is transcriptionally upregulated by heat shock, calcium and heavy metals. Mol Immunol 1996, 33:379-386. 27. Dreher D, Vargas JR, Hochstrasser DF, Junod AF: Effects of oxidative stress and Ca2+ agonists on molecular chaperones in human umbilical vein endothelial cells. Electrophoresis 1995, 16:1205-1214. 28. Conway EM, Liu L, Nowakowski B, Steiner-Mosonyi M, Ribeiro SP, Michalak M: Heat shock-sensitive expression of calreticulin. In vitro and in vivo up-regulation. J Biol Chem 1995, 270:17011-17016. 29. Kuwabara K, Pinsky DJ, Schmidt AM, Benedict C, Brett J, Ogawa S, Broekman MJ, Marcus AJ, Sciacca RR, Michalak M, Wang F, Pan YC, Grunfeld S, Patton S, Malinski T, Stern DM, Ryan J: Calreticulin, an antithrombotic agent which binds to vitamin K-dependent coagulation factors, stimulates endothelial nitric oxide production, and limits thrombosis in canine coronary arteries. J Biol Chem 1995, 270:8179-8187. 30. Karska K, Tuckova L, Steiner L, Tlaskalova-Hogenova H, Michalak M: Calreticulin--the potential autoantigen in celiac disease. Biochem Biophys Res Commun 1995, 209:597-605. 31. Boehm J, Orth T, Van Nguyen P, S=F6ling H-D: Systemic lupus erythematosus is associated with increased auto-antibody titers against calreticulin and grp94, but calreticulin is not the Ro/SS-A antigen. Eur J Clin Invest 1994, 24:248-257. 32. Zhu J, Newkirk MM: Viral induction of the human autoantigen calreticulin. Clin Invest Med 1994, 17:196-205. 33. Ben-Chetrit E: The molecular basis of the SSA/Ro antigens and the clinical significance of their autoantibodies. Br J Rheumatol 1993, 32:396-402. 34. McCauliffe DP, Sontheimer RD: Molecular characterization of the Ro/SS-A autoantigens. J Invest Dermatol 1993, 100:73S-79S. 35. Atreya CD, Singh NK, Nakhasi HL: The rubella virus RNA binding activity of human calreticulin is localized to the N-terminal domain. J Virol 1995, 69:3848-3851. 36. Singh NK, Atreya CD, Nakhasi HL: Identification of calreticulin as a rubella virus RNA binding protein. Proc Natl Acad Sci USA 1994, 91:12770-12774. 37. Dupuis M, Schaerer E, Krause K-H, Tschopp J: The calcium-binding protein calreticulin is a major constituent of lytic granules in cytolytic T lymphocytes. J Exp Med 1993, 177:1-7. 38. Stendahl O, Krause K-H, Krischer J, Jerstrom P, Theler JM, Clark RA, Carpentier JL, Lew DP: Redistribution of intracellular Ca2+ stores during phagocytosis in human neutrophils. Science 1994, 265:1439-1441. 39. Nakamura M, Moriya M, Baba T, Michikawa Y, Yamanobe T, Arai K, Okinaga S, Kobayashi T: An endoplasmic reticulum protein, calreticulin, is transported into the acrosome of rat sperm. Exp Cell Res 1993, 205:101-110. 40. Dedhar S: Novel functions for calreticulin: Interaction with integrins and modulation of gene expression. Trends Biochem Sci 1994, 19:269-271. 41. White TK, Zhu Q, Tanzer ML: Cell surface calreticulin is a putative mannoside lectin which triggers mouse melanoma cell spreading. J Biol Chem 1995, 270:15926-15929. 42. Gray AJ, Park PW, Broekelmann TJ, Laurent GJ, Reeves JT, Stenmark KR, Mecham RP: The mitogenic effects of the B chain of fibrinogen are mediated through cell surface calreticulin. J Biol Chem 1995, 270:26602-26606. 43. Krause K-H, Michalak M: Calreticulin. Cell 1997, 88:439-443. Dr. Michal Opas Department of Anatomy & Cell Biology University of Toronto 1 King's College Circle Medical Sciences Building Toronto, Ontario, M5S 1A8 Canada phone: (416) 978-8947 fax: (416) 978-3954 e-mail: m.opas@utoronto.ca www homepage: http://www.utoronto.ca/anatomy/opas/start.htm From owner-repertoires@net.bio.net Tue Jul 22 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Stephen Donoghue Newsgroups: bionet.molecules.repertoires Subject: Re: Pharmacia kit Date: 23 Jul 1997 00:55:45 -0700 Organization: Pathology Department , Leicester, U.K. Lines: 16 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <33D342E5.6BFD@le.ac.uk> References: <01IL4JMH19228Y665C@ubaclu.unibas.ch> NNTP-Posting-Host: net.bio.net I am about to embark on the phage display ship and i was wondering if anyone could recommend the Pharmacia kit. I will be amplifying the V genes from both a spleen repertoire and established monoclonal cell lines. I've bought the ScFv module kit and i was wondering if anyone knows the primer sequences for the light and heavy primer mixes. Cheers Stephen Donoghue Robert Kilpatrick Clinical Sciences Building Leicester Royal Infirmary, Leicester Le2 7LX U.K. email sed7@le.ac.uk From owner-repertoires@net.bio.net Wed Jul 23 23:00:00 1997 Path: biosci!biosci!not-for-mail From: "Dimitris K. Agrafiotis" Newsgroups: bionet.molecules.repertoires Subject: preprints available Date: 24 Jul 1997 02:31:14 -0700 Organization: 3-Dimensional Pharmaceuticals, Inc. Lines: 18 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <33D669AE.F8BF8EB0@3dp.com> Reply-To: "Dimitris K. Agrafiotis" NNTP-Posting-Host: net.bio.net Preprints from some of our publications and reviews can now be found on our web site under http://www.3dp.com/papers.html, in gziped PostScript format. Please let me know if you have any difficulty accessing or reading these files. Those of you who received my last response without the attachment, please visit this site and download the manuscript(s) of interest. Thanks, -- Dimitris K. Agrafiotis, PhD | E-mail: dimitris@3dp.com 3-Dimensional Pharmaceuticals, Inc. | Tel: (610) 458-6045 665 Stockton Drive, Suite 104 | Fax: (610) 458-8249 Exton, PA 19341 From owner-repertoires@net.bio.net Wed Jul 23 23:00:00 1997 Path: biosci!biosci!not-for-mail From: M.Foley@latrobe.edu.au (Dr Michael Foley) Newsgroups: bionet.molecules.repertoires Subject: deletions in phage peptides Date: 24 Jul 1997 02:33:04 -0700 Organization: La Trobe University Lines: 17 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <5r6evc$nm4$1@news.latrobe.edu.au> NNTP-Posting-Host: net.bio.net Hi All, We have recently been screening random phage peptide libraries with 15 amino acid inserts (from George Smith and Jamie Scotts labs) and have noticed a couple of clones coming through with deletions. One clone from one library had 3 amino acids deleted close to the N-terminus and another clone from the other library had 13 amino acids deleted. I have read a few accounts of this happening, but I was wondering what other peoples experiences were. Cheers, Mick From owner-repertoires@net.bio.net Thu Jul 24 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Brian Walker Newsgroups: bionet.molecules.repertoires Subject: UK Lectureship in Molecular Cell Biology Date: 25 Jul 1997 02:07:38 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 34 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <5r9q2u$qat@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net LECTURESHIP IN MOLECULAR CELL BIOLOGY The Queen's University of Belfast School of Biology and Biochemistry Centre for Peptide and Protein Engineering Applications are invited from individuals with an excellent research record in molecular cell biology and, preferably, proven ability in teaching, for the above mentioned post. Applicants must have an honours degree (or equivalent) and a Ph.D. in Biochemistry, Molecular Biology, or a closely related subject. Experience of applying molecular biological techniques to the study of one of the following: signal transduction pathways, molecular processing, intracellular proteolysis, or mode of action of transcription factors, is essential. Expertise in differential display and yeast two-hybrid systems, would also be highly desirable. The successful candidate will be expected to develop an independent research programme and to foster collaborative links with other members of the Centre for Peptide and Protein Engineering, to which he/she will be associated. The successful candidate must also be committed to research-led teaching and will be expected to make an immediate (although relatively light) contribution to undergraduate teaching. Informal enquiries may be directed to Dr. B. Walker, School of Biology and Biochemistry, telephone +44 (0)1232 272047, FAX +44 (0)1232 236505, Email: brian.walker@qub.ac.uk. Committed to an Equal Opportunities policy and selection on merit, the University welcomes applications from all sections of the community. Under its affirmative action programme, the University particularly welcomes applications from women for academic posts. From owner-repertoires@net.bio.net Sun Jul 27 23:00:00 1997 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.molecules.repertoires Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 28 Jul 1997 02:08:21 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 236 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <199707280900.CAA00635@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 30-JUL-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 0) BIOSCI NEEDS YOUR SUPPORT!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- computer daresbury.ac.uk (also known as dl.ac.uk): ------------------------------------------------- To subscribe and unsubscribe to/from the BIOSCI lists, you need to specify the full USENET newsgroup name with "bionet-news." prepended. The USENET newsgroup names are listed in the BIOSCI Information sheet on the Web at http://www.bio.net/. For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. From owner-repertoires@net.bio.net Wed Jul 30 23:00:00 1997 Path: biosci!biosci!not-for-mail From: rakonj@rockvax.rockefeller.edu (Jasna Rakonjac) Newsgroups: bionet.molecules.repertoires Subject: purification of phage Date: 31 Jul 1997 10:04:49 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 43 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net >Dear all, > >I'm currently displaying my favorite protein on pIII using a modified pHEN1 >phagemid. I would like to test these fusion-phage for enzymatic activity, >therefore I need them as pure as possible. I know that CsCl-centrifugation is >a possibility, but I really dont like the idea of exposing my protein to >such high amounts of salt. Thus I wondered if any of you know of a dialysis >membrane with an extreme high cut-off that could be used for the purpose? > >>From SDS-PAGE of precipitated phage it appears that 'junk' proteins co-ppt, >have any of you tried to get rid of these by e.g. dialysis or gel-filtration? >If so, I would appreciate to hear from you! > >Thanks in advance, >Troels Wind >Aarhus University >Denmark Hi Troels, You can purify phage using Centricon filters (manufactured by Amicon). You can use either the big units for filtration by pressure, or the smaller that use centrifuge force. They come in the cutoff sizes of 300 000 and 500 000 daltons. In my experience, big units are better. You can let all the medium be filtered out, and then resuspend the phage by placing the filter into your buffer of choice for several hours at 4degC.These phage are not, however, as clean as CsCl purified ones. Cheers, Jasna Jasna Rakonjac Zinder-Model Laboratory The Rockefeller University 1230 York Avenue, Box 24 New York, NY 10021 U.S.A. Phone: 212-327-8649 Fax: 212-327-7850 From owner-repertoires@net.bio.net Wed Jul 30 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Troels Wind Newsgroups: bionet.molecules.repertoires Subject: Purification of phage by dialysis Date: 31 Jul 1997 03:50:19 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 20 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <5rpo4c$9jt@mserv1.dl.ac.uk> NNTP-Posting-Host: net.bio.net Dear all, I'm currently displaying my favorite protein on pIII using a modified pHEN1 phagemid. I would like to test these fusion-phage for enzymatic activity, therefore I need them as pure as possible. I know that CsCl-centrifugation is a possibility, but I really dont like the idea of exposing my protein to such high amounts of salt. Thus I wondered if any of you know of a dialysis membrane with an extreme high cut-off that could be used for the purpose? >From SDS-PAGE of precipitated phage it appears that 'junk' proteins co-ppt, have any of you tried to get rid of these by e.g. dialysis or gel-filtration? If so, I would appreciate to hear from you! Thanks in advance, Troels Wind Aarhus University Denmark From owner-repertoires@net.bio.net Thu Jul 31 23:00:00 1997 Path: biosci!biosci!not-for-mail From: M.Foley@latrobe.edu.au (Dr Michael Foley) Newsgroups: bionet.molecules.repertoires Subject: cysteine constraints Date: 1 Aug 1997 02:42:48 -0700 Organization: La Trobe University Lines: 34 Sender: daemon@net.bio.net Approved: a.wallace@qub.ac.uk Distribution: world Message-ID: <5rrhl1$oo1$1@news.latrobe.edu.au> NNTP-Posting-Host: net.bio.net Hi all, First of all a week or so ago I asked this: >We have recently been screening random phage peptide libraries with 15 amino >acid inserts (from George Smith and Jamie Scotts labs) and have noticed a >couple of clones coming through with deletions. One clone from one library had >3 amino acids deleted close to the N-terminus and another clone from the other >library had 13 amino acids deleted. I have read a few accounts of this >happening, but I was wondering what other peoples experiences were. Thanks to Richard Odessey and James Burritt and others for the replys. It looks like deletions are common and can sometimes lead to useful sequences. This is what we have seen. We have seen a three amino acid deletion and a 13amino acid deletion from a 15 mer library. The former binds specifically to our Ab the latter doesn't. I have another question if you don't mind. Has anyone tried to incubate phage containing cysteine constraints in the peptide with beta mercaptoethanol or DTT and examine the effects on binding. (obviously they have to be binding phage to begin with). Do you have to synthesise the peptide to do this or can you test it directly on the phage displayed peptide? Will these reagents stuff up the phage? Any ideas welcome. Cheers, Mick