From owner-7tms_r@net.bio.net Sat Nov 07 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Ian Murray" Newsgroups: bionet.molbio.proteins.7tms_r Subject: Receptor isolation: Crosslinking, Ligand blot, Ligand affinity column Date: 8 Nov 1998 18:52:54 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <725le6$o0@net.bio.net> NNTP-Posting-Host: net.bio.net Hello, I posted the following message on the group and received helpful information concerning GPCR and ligand blots. Message-Id: <199712202156.NAA15805@net.bio.net> I was wondering if I could receive clarification on a matter. Apparently there has been no successful method of receptor isolation by these methods ... I was unclear if this was referring to BOTH ligand blots and crosslinking OR just only ligand blots?. 1. We have tried both Crosslinking and ligand blots. Crosslinking yields very small amounts of receptor, and is apparently not very efficient. Ligand blots only work for a select few ligands, where binding occurs on the N terminal loops. 2. So, ligand affinity columns??? Are they better. We have read that they yield larger amounts of protein. Does anyone have experience in this area and could be of assistance?? I would greatly appreciate your help, Ivan Murray From owner-7tms_r@net.bio.net Sat Nov 14 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Brian Wickes Newsgroups: bionet.molbio.proteins.7tms_r Subject: Post-Doc Date: 15 Nov 1998 09:22:57 -0800 Organization: University of Texas Health Science Center Lines: 28 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <72n2lh$mvc@net.bio.net> NNTP-Posting-Host: net.bio.net POST-DOCTORAL POSITION IN MEDICAL MYCOLOGY A Post-Doctoral position is available for studying the role of mating-type associated genes in virulence of Cryptococcus neoformans, an important human fungal pathogen. Projects include the isolation and characterization of genes involved in mating, hyphal development, and/or virulence. Experience in molecular biology is preferred. The position is located in the Dept. of Microbiology at the University of Texas Health Science Center at San Antonio (UTHSCSA) and is funded for up to three years. The Health Science Center has one of the largest groups of internationally recognized mycologists working on human fungal pathogens and offers an excellent opportunity to enter the field of medical mycology. UTHSCSA is an affirmative action/equal opportunity employer. Salary is highly competitive. Applications may be sent by mail, fax, or e-mail and must include the following: a detailed C.V. and contact information for three references. Send to: Brian Wickes, Ph.D. Dept. of Microbiology, Rm 5.023V 7703 Floyd Curl Dr. University of Texas Health Science Center at San Antonio San Antonio, TX 78284-7758 Tel (210) 567-3938 Fax (210) 567-6612 e-mail wickes@uthscsa.edu From owner-7tms_r@net.bio.net Sat Nov 14 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Athan Kuliopulos Newsgroups: bionet.molbio.proteins.7tms_r Subject: Signal Transduction and Cancer Postdoc Position Avail-Boston Date: 15 Nov 1998 10:24:38 -0800 Organization: NEMC Lines: 56 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <72n696$35i@net.bio.net> Reply-To: "Kuliopoulos@HemOnc"@NEMC.nemc.org NNTP-Posting-Host: net.bio.net Postdoctoral Position Available Molecular Cardiology Research Institute Tufts University School of Medicine-NEMC Boston, MA We have an immediate opening for a Postdoctoral Fellow to study the role of the thrombin receptor in cancer. In addition to its critical roles in hemostasis and thrombosis, the thrombin receptor is likely to play a major part in vascularization of tissue during embryogenesis and malignant processes. The PAR1 thrombin receptor serves as the paradigm for a rapidly developing field of protease-activated receptors which are all activated by cleavage at a specific peptide bond by their cognate protease. The thrombin-cleaved N-terminus of PAR1, S42FLLRN47, acts as an intramolecular ligand that binds to the body of the receptor by a poorly understood mechanism. Peptides corresponding to the N-terminus, such as SFLLRN, can elicit a subset of the PAR1 responses independent of receptor proteolysis. In addition to platelets and endothelial cells, PAR1 has been identified in fibroblasts, smooth muscle cells, macrophages, lymphocytes, and neurons. Upon intra- or inter-molecular liganding, the signal is transduced through Gq, Gai, and/or G12/13 that leads to stimulation of various signaling pathways that include adenylate cyclase, phospholipase C-b (PLC-b), phospholipase A2, protein kinase C, PI3-kinase-g, and MAP kinases. Three additional protease-activated receptors have recently been identified, PAR2, PAR3, and PAR4 but their respective roles are nearly completely unknown. We have made the intriguing observation that high levels of PAR1 expression and activity occur in a non-small cell lung epithelial cancer cell line. Interestingly, a recent report suggests the involvement of the PAR1 receptor in cellular invasion associated with tumor progression. The goals of this project include extending this preliminary observation to see whether other cancer cell lines including primary breast carcinoma cell lines will demonstrate a direct correlation between the level of expression and activity of PAR receptors and invasiveness. Our laboratory is located within the Molecular Cardiology Research Institute, a modern, state-of-the-art facility supported by several core units including automated DNA sequencing, oligonucleotide and peptide synthesis, plasmid preparation, small animal surgical and physiology lab, flow cytometry, and transgenic mouse facility. Qualifications for this position are a Ph.D. degree and experience in mammalian cell transfection and expression. Interested candidates should e-mail a description of their research interests, a CV, and names of three references to: Athan Kuliopulos, MD., Ph.D. Assistant Professor Division of Hematology-Oncology Tufts-NEMC Box 832 750 Washington Street Boston, MA 02111 617-636-8482 617-636-4833 (fax) akuliopu@opal.tufts.edu