Adding membrane signal to cytoplasmic protein

Michael Coady coady at ERE.UMontreal.CA
Thu Oct 27 16:27:49 EST 1994


In article <1994Oct25.210606.8762 at mbcf> mhughes at mbcf.stjude.org writes:
>In article <01HIIC3VA6HU0008KV at UNCVX1.OIT.UNC.EDU>, ORSULIC at UNCVX1.OIT.UNC.EDU writes:
>> I would like to generate the form of the protein that would be tethered to the
>> membrane. Any suggestions, protocols or references of successful addition of a
>> myristilation or membrane signal to other proteins would be appreciated. 
>	You might want to try adding a ras-like CAAX sequence to the c-terminus
>of you protein.  I don't know if this has been done successfully before or not, 
>but I'm almost positive that this sequence alone comprizes the entire
>myristilation signal for ras and many other proteins, and may work for
>armadillo.  

	Another possibility is described in Analytical Biochemistry, vol
217, pp 333-335 (1994).  The authors wanted to express luciferase as a
membrane protein so they fused the cDNA to that of the part of the 
HT1a receptor.  They found that their fusion was enzymatically active
and associated with the membranous fraction of the cell homogenate.
The authors are also kindly making the vector available to those
(like myself) who wanted it.  'Fraid I haven't tried it yet with
my fusion protein so I can't add any more to this thread.

Mike

-- 
Michael J. Coady
COADY at ERE.UMONTREAL.CA
The opinions expressed above are solely those of the author and do not,
in any way, shape or form, represent the Universite de Montreal.



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