ras p21 and other Gn proteins are detected in mammalian cell lines by [gamma-35S]GTP gamma S binding.
The presence of guanine nucleotide binding proteins in mouse and human cell lines was investigated using [gamma-35S]GTP gamma S and [gamma-32P]GTP. Cell lysate polypeptides were separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis and transferred to nitrocellulose. Incubation of the nitrocellulose blots with [gamma-35S]GTP gamma S identified 9 distinct GTP-binding polypeptides in all lysates. One of these is the ras oncogene product, p21, as demonstrated by subsequent immunochemical staining of the nitrocellulose blots. We have shown that this procedure provides a sensitive method for detection of p21 in culture cell lines.
Comerford
J G
JG
School of Biological Sciences, University of East Anglia, Norwich, England.
Gibson
J R
JR
Dawson
A P
AP
Gibson
I
I
D000818:Animals; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D019204:GTP-Binding Proteins / Q000032:analysis* / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D011518:Proto-Oncogene Proteins / Q000032:analysis* / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras); D011865:Radioisotope Dilution Technique; D013462:Sulfur Radioisotopes; D013873:Thionucleotides / Q000378:metabolism*
D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D013462:Sulfur Radioisotopes; D013873:Thionucleotides; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)