The submembranous location of p11 and its interaction with the p36 substrate of pp60 src kinase in situ.
p36, a major cytoplasmic substrate of pp60 src kinase, is present beneath the plasma membrane. It can be isolated either as a monomer or as a heterotetramer (protein I) containing two copies each of p36 and a unique p11 polypeptide. To compare the expression rules of p36 and p11 as well as their cellular distributions, monoclonal antibodies to the two porcine proteins were isolated. In tissue culture cells p11-specific antibodies decorated the same submembranous compartment previously seen with antibodies to p36 and fodrin or spectrin and followed the p36 images under all fixation/extraction conditions tested. Immunofluorescence microscopy on tissue sections showed coincident expression patterns of both proteins confirming and extending previous results with p36 antibodies. Antibodies with limited cross-species reaction have been used to trace the fate of porcine p11 and p36 injected into cultured cells. Both proteins are incorporated in the submembranous compartment, where they remain in Triton cytoskeletons prepared in the presence but not in the absence of Ca2+. The incorporation of p36 in vivo conforms with its Ca2+-dependent binding to actin, fodrin, and certain phospholipids in vitro. In contrast, the incorporation of p11 seems to depend on an in situ interaction with p36 or an exchange with endogenous p11 present on p36. The combined results indicate a strong coupling of p11 and p36 in cellular compartmentalization and tissue differentiation.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002417:Cattle; D002451:Cell Compartmentation; D002460:Cell Line; D011232:Chemical Precipitation; D003593:Cytoplasm / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D015688:Oncogene Protein pp60(v-src); D010750:Phosphoproteins / Q000276:immunology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D012191:Retroviridae Proteins / Q000378:metabolism*; D009418:S100 Proteins / Q000276:immunology; D013552:Swine
D000911:Antibodies, Monoclonal; D010750:Phosphoproteins; D012191:Retroviridae Proteins; D009418:S100 Proteins; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src)