Phosphorylation reduces the affinity of protein 4.1 for spectrin.
The phosphorylation of protein 4.1 by the membrane kinase and casein kinase A has been investigated. Each of these kinases catalyzed the incorporation of 2 mol of phosphate per mole of protein 4.1. The presence of both kinases in the reaction mixture did not lead to an increase in the incorporation of phosphates into the protein. An analysis of the acid hydrolysis products of the 32P-labeled protein 4.1 indicated that the radioactivities were distributed between phosphothreonine and phosphoserine in a ratio of about 2 to 1. The effects of phosphorylation on the binding of protein 4.1 to spectrin were investigated by using sucrose density gradient centrifugation. The affinity of protein 4.1 for spectrin was reduced about 5-fold, from a KD of 2 X 10(-6) M to a KD of 9.4 X 10(-6) M, by phosphorylation. The phosphorylation of spectrin, on the other hand, appeared to increase slightly its affinity for protein 4.1. The results suggest that phosphorylation may lead to a relaxation of the cytoskeletal network and the formation of a more flexible membrane structure that is important to red cell function.
P S
PS
Eder
C J
CJ
Soong
M
M
Tao
D000255:Adenosine Triphosphate / Q000097:blood; D000596:Amino Acids / Q000032:analysis; D001798:Blood Proteins / Q000302:isolation & purification / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins; D009479:Neuropeptides; D010750:Phosphoproteins / Q000097:blood / Q000302:isolation & purification; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011485:Protein Binding; D013049:Spectrin / Q000302:isolation & purification / Q000378:metabolism*
D000596:Amino Acids; D001798:Blood Proteins; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D009479:Neuropeptides; D010750:Phosphoproteins; D010761:Phosphorus Radioisotopes; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; D013049:Spectrin; D000255:Adenosine Triphosphate