Identification of a protein cross-reacting with anti-phosphotyrosine antibodies in yeast insoluble cytoplasmic matrices.
Immunoblot analysis with anti-phosphotyrosine antibodies of total extracts from exponentially growing yeast cells reveals a unique cross-reactive polypeptide of about 75 Kd (p75). The specificity of the immunodecorations has been checked by experiments of competition with phosphoaminoacids. A common behaviour has been observed for the 75 kd band and the 170 kd band corresponding to the platelet-derived growth factor receptor from Swiss 3T3 cells, which it has been known to be autophosphorylated on tyrosine upon ligand binding and used as a control throughout this work. We have found that p75 is associated to detergent insoluble cytoplasmic matrices. The stability of p75 detection by antibodies following treatments that specifically hydrolyze phosphohistidine and its susceptibility to potato acid phosphatase treatment provide further evidences that the epitope recognized by these antibodies in the yeast p75 polypeptide is indeed phosphotyrosine.
Grandori
R
R
Dept. of General Physiology and Biochemistry, University of Milano, Italy.
Vai
M
M
Di Renzo
M F
MF
Alberghina
L
L
Popolo
L
L
D000908:Antibodies, Fungal / Q000032:analysis*; D000918:Antibody Specificity; D002458:Cell Fractionation; D003429:Cross Reactions; D003593:Cytoplasm / Q000032:analysis*; D005656:Fungal Proteins / Q000032:analysis* / Q000276:immunology; D008970:Molecular Weight; D017830:Octoxynol; D019000:Phosphotyrosine; D011092:Polyethylene Glycols; D012441:Saccharomyces cerevisiae / Q000032:analysis* / Q000166:cytology; D012995:Solubility; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology
D000908:Antibodies, Fungal; D005656:Fungal Proteins; D019000:Phosphotyrosine; D011092:Polyethylene Glycols; D014443:Tyrosine; D017830:Octoxynol