Isolation of a major human placental substrate for the epidermal growth factor (urogastrone) receptor kinase: immunological cross-reactivity with transducin and sequence homology with lipocortin.
Using as a starting material either a detergent extract or a protein fraction eluted from membranes with ethylene glycol bis (beta-aminoethyl ether)-N,N'-tetraacetic acid, we have isolated from human placental membranes a major substrate for the epidermal growth factor (urogastrone) receptor kinase (EGF kinase). The substrate was isolated both in an intact form, having a molecular mass of approximately 38-kDa (p38), and in a 35-kDa form (p35) representing a proteolytic cleavage product of p38. Both p38 and p35 cross-reacted with antibodies directed against bovine retinal transducin, but did not cross-react with antibodies directed against the 35-kDa beta subunit of human placental G-protein. Antisera directed against the placental EGF kinase substrate failed to react with either bovine or human placental src kinase substrate, p36. Conversely, antisera directed against p36 reacted only poorly with placental p38 or p35. Although p38 had a blocked amino terminus that precluded sequence analysis, p35 yielded an N-terminal sequence that was identical with residues 13-36 of human lipocortin. Our data clearly distinguish p38 from the previously described intestinal calcium binding protein calpactin I or p36 that is also a tyrosine kinase substrate, and our work points to a close relationship (if not identity) between p35 and a 35-kDa EGF receptor kinase substrate previously characterized in A431 cells. We conclude that p38 and p35, which very likely represent human placental lipocortin, may share only limited epitope homology with transducin alpha subunit; however, the possibility that p38, along with intestinal p36 and with a family of related calcium binding proteins, may, like transducin, play a role in receptor-mediated transmembrane signaling is discussed.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D017302:Annexins; D003429:Cross Reactions; D066246:ErbB Receptors / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000032:analysis*; D008566:Membranes / Q000032:analysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D010920:Placenta / Q000032:analysis*; D011494:Protein Kinases / Q000378:metabolism*; D014020:Tissue Extracts / Q000032:analysis* / Q000378:metabolism; D015293:Transducin
D000596:Amino Acids; D017302:Annexins; D006023:Glycoproteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D014020:Tissue Extracts; D011494:Protein Kinases; D066246:ErbB Receptors; D015293:Transducin