The ezrin-like family of tyrosine kinase substrates: receptor-specific pattern of tyrosine phosphorylation and relationship to malignant transformation.
A method for the isolation of tyrosine kinases substrates was developed. The method takes advantage of immuno-affinity purification of an entire set of proteins phosphorylated by tyrosine kinases, followed by generation of antisera against the purified protein pool and immunological screening of bacterial expression libraries with these antisera. By applying this methodology to the study of the phosphorylation events triggered by activation of the epidermal growth factor receptors, we have isolated several cDNAs encoding novel putative tyrosine kinase substrates. One of these cDNAs encodes radixin, a protein belonging to the band 4.1 family of proteins and highly related to ezrin and moesin. We demonstrated that, despite a high degree of relatedness, these three proteins exhibit a distinct receptor-specific pattern of phosphorylation, raising the possibility that they might mediate receptor-specific cellular changes. In addition the generation of antibodies specific for either radixin, ezrin or moesin allowed us to show that a previously described tumor transplantation antigen is indeed ezrin, thus implicating this protein in the determination of the biological phenotype of certain tumors.
Fazioli
F
F
Laboratory of Molecular and Cellular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Wong
W T
WT
Ullrich
S J
SJ
Sakaguchi
K
K
Appella
E
E
Di Fiore
P P
PP
D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis; D001483:Base Sequence; D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism; D002471:Cell Transformation, Neoplastic; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins; D066246:ErbB Receptors / Q000378:metabolism; D006649:Histocompatibility Antigens / Q000032:analysis; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D009479:Neuropeptides; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D011817:Rabbits; D017479:Receptors, Platelet-Derived Growth Factor / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*; D000099149:Ezrin
D000951:Antigens, Neoplasm; D001798:Blood Proteins; D003598:Cytoskeletal Proteins; D066246:ErbB Receptors; D006649:Histocompatibility Antigens; D008565:Membrane Proteins; D008840:Microfilament Proteins; D009479:Neuropeptides; D010750:Phosphoproteins; D011505:Protein-Tyrosine Kinases; D011506:Proteins; D017479:Receptors, Platelet-Derived Growth Factor; D014443:Tyrosine; D000099149:Ezrin; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; C029237:tumor-associated transplantation antigen; C071058:moesin; C071776:radixin