Phosphorylation and dephosphorylation of spectrin from human erythrocyte ghosts under physiological conditions: autocatalysis rather than reaction with separate kinase and phosphatase.
The mechanism of phosphosylation and dephosphorylation of spectrin from human erythrocyte membranes has been examined under closely physiological conditions. The results support the hypothesis that spectrin is an autophosphorylating and dephosphorylating system. (i) Extraction from ghosts of up to 85% of the kinase (casein kinase) suggested to catalyze the reaction [see Fairbanks, G., Avruch, J., Dino, E. J. & Patel, V. P. (1978) J. Supramol. Struct. 9, 97--112] only slightly reduced spectrin component 2 phosphorylation and did not affect ATP-induced changes in the ghosts' shapes. (ii) A spectrin--actin complex isolated from endocytotic inside-out vesicles under hyperteonic conditions contained virtually no casein kinase activity and still exhibited a largely intact phosphorylation machinery. (iii) Photoaffinity labeling experiments indicated that spectrin component 2 fulfills the necessary prerequisite of the hypothesis--i.e., it contains its own ATP-binding site. (iv) Under various conditions, spectrin phosphorylation and dephospohrylation seem to be tightly coupled. The implications of these findings for the understanding of spectrin function and the maintenance of erythrocyte shape are discussed.
D000199:Actins / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000345:Affinity Labels / Q000378:metabolism; D002364:Caseins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000201:enzymology*; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism*; D008954:Models, Biological; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000032:analysis; D012965:Sodium Chloride / Q000494:pharmacology; D013049:Spectrin / Q000378:metabolism* / Q000528:radiation effects; D014466:Ultraviolet Rays
D000199:Actins; D000345:Affinity Labels; D002364:Caseins; D008565:Membrane Proteins; D047428:Protein Kinase Inhibitors; D013049:Spectrin; D012965:Sodium Chloride; D000255:Adenosine Triphosphate; D011494:Protein Kinases