cell wall permeability

navin navin
Tue Sep 26 20:32:19 EST 1995


Hi,

        In order to map the binding site(s) on a receptor for a peptide hormone
I'm studying, I want to make a library of semi-random point-mutants, express
this library of mutants in yeast, and then screen by FACS (using a fluorescent
derivative of the hormone as a probe) for mutants that don't bind to the
hormone. Our lab does not have mammalian tissue culture facilities (which is
why we want to do our studies in yeast), but there is a departmental FACS.
        How permeable is the yeast cell wall? I realize that for
immunofluoresence,the cell wall must be removed. But the peptides of interest
are only 30-50 residues long (an order of magnitude smaller than antibodies).
Would these have a problem getting through the cell wall to the plasma membrane
(where the receptor would be localized)?

Thanks,

Navin Pokala
navin at uclink4.berkeley.edu




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