Purification of band 7.2b, a 31-kDa integral phosphoprotein absent in hereditary stomatocytosis.
A 31-kDa human erythrocyte integral protein, band 7.2b, has been purified to better than 95% homogeneity. The polypeptide was found to be insoluble in most detergents and was isolated in denatured form by gel filtration in the presence of sodium dodecyl sulfate and preparative electrophoresis. In intact erythrocytes that were equilibrated with 32Pi, band 7.2b was phosphorylated in response to exogenous dibutyryl cAMP. The peptide is also palmitylated, as shown by its incorporation of radioactivity when intact erythrocytes were incubated with [9,10-3H]palmitic acid. Antisera to band 7.2b were raised in rabbits, and these antibodies cross-react with 31-kDa polypeptides in human liver and kidney. Immunoblots of red cells from a number of other species were negative, with the exception of a cross-reacting 23-kDa polypeptide in rat erythrocyte membranes. Band 7.2b was absent in erythrocyte membranes from an individual with overhydrated hereditary stomatocytes.
Wang
D
D
Department of Biochemistry, Wayne Street University, Detroit, Michigan 48201.
Mentzer
W C
WC
Cameron
T
T
Johnson
R M
RM
D000596:Amino Acids / Q000032:analysis; D000745:Anemia, Hemolytic, Congenital / Q000378:metabolism*; D001345:Autoradiography; D001798:Blood Proteins / Q000302:isolation & purification*; D015153:Blotting, Western; D003994:Bucladesine / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000187:drug effects; D006801:Humans; D008565:Membrane Proteins / Q000302:isolation & purification*; D010750:Phosphoproteins / Q000302:isolation & purification*; D010766:Phosphorylation
D000596:Amino Acids; D001798:Blood Proteins; D008565:Membrane Proteins; D010750:Phosphoproteins; D003994:Bucladesine