Possible binding proteins of ras p21 in human erythrocyte membrane.
The direct binding protein(s) of ras p21 was investigated in the inside-out vesicles of human erythrocyte ghosts using the pure v-Kirsten (Ki)-ras p21 synthesized in E. coli. The bound ras p21 was detected immunochemically using an anti-v-Ki-ras p21 monoclonal antibody. ras p21 was overlaid on the vesicle proteins immobilized on a nitrocellulose sheet transferred from the gel of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. ras p21 bound to bands 4.2 and 6. ras p21-binding to bands 4.2 and 6 was reduced by prior incubation of ras p21 with the purified band 4.2 or 6 protein. Furthermore, when ras p21 was mixed with inside-out vesicles and then centrifuged, ras p21 was coprecipitated with the vesicles. Prior digestion of the vesicles with trypsin reduced this binding significantly. These results indicate that v-Ki-ras p21 can bind directly to bands 4.2 and 6 of human erythrocyte membranes as fat as tested in an in vitro cell-free system.
Tanimoto
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T
Department of Biochemistry, Kobe University School of Medicine.
D001798:Blood Proteins / Q000378:metabolism*; D002352:Carrier Proteins / Q000097:blood*; D003598:Cytoskeletal Proteins; D004910:Erythrocyte Membrane / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008565:Membrane Proteins; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D001798:Blood Proteins; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; C054632:band 6 protein, human; C044791:erythrocyte membrane band 4.2 protein; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D014357:Trypsin; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)