protein-protein interactions

NANTEL at UNCVX1.BITNET NANTEL at UNCVX1.BITNET
Tue Aug 25 07:17:00 EST 1992


I personnaly know (and use) two other techniques for studying protein-
protein interactions on top of crosslinking and immunoprecipitation:

Far western blot: Similar to a western blot except that a labelled protein
is used as a probe. The probe can be biotinylated or labelled with 32P,
125I or 35S. The following references will be useful:
        Oncogene (1990), 5, 451-458
        Mol Endocrinol (1991), 5, 256-266
        Genes & Dev (1992), 6, 439-453
        Science (1992), 257, 803-805

Affinity chromatography: Express one of your protein as a GST (or MBP)
fusion in E. coli. These fusion proteins will then bind to gluthatione
sepharose or amylose resins and can be used to fish out binding proteins.
Numerous labs have used this technique which was originally described by
Kaelin et al. in Cell (1991), 64, 521-532.

Hope these can be of help. Sorry I didn't answer you directely but something
in your address make my computer gag.

Andre Nantel
Dept. of biology
University of North Carolina-Chapel Hill
"Murphy's law is strong in molecular biology"



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