[Isolation and properties of three forms of phosphorylase and phosphorylase kinase from human skeletal muscle].
Three forms of phosphorylase (I, II and III), two of which (I and II) were active in the presence of AMP and one (III) was active without AMP, were isolated from human skeletal muscles. The pI values for phosphorylases b(I) and b(II) were found to be identical (5.8-5.9). During chromatofocusing a low molecular weight protein (M(r) = 20-21 kDa, pI 4.8) was separated from phosphorylase b(II). This process was accompanied by an increase of the enzyme specific activity followed by its decline. During reconstitution of the complex the activity of phosphorylase b(II) returned to the initial level. Upon phosphorylation the amount of 32P incorporated into phosphorylase b(II) was 2 times as low as compared with rabbit phosphorylase b and human phosphorylase b(I). It may be supposed that in the human phosphorylase b(II) molecule one of the two subunits undergoes phosphorylation in vivo. This form of the enzyme is characterized by a greater affinity for glycogen and a lower sensitivity to allosteric effectors (AMP, glucose-6-phosphate, caffeine) compared with phosphorylase b(I). Thus, among the three phosphorylase forms obtained in this study, form b(II) is the most unusual one, since it is partly phosphorylated by phosphorylase kinase to form a complex with a low molecular weight protein which stabilizes its activity. A partially purified preparation of phosphorylase kinase was isolated from human skeletal muscles. The enzyme activity necessitates Ca2+ (c0.5 = 0.63 microM). At pH 6.8 the enzyme is activated by calmodulin (c0.5 = 15 microM). The enzyme activity ratio at pH 6.8/8.2 is equal to 0.18.
Skolysheva
L K
LK
Shur
S A
SA
Vul'fson
P L
PL
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D002853:Chromatography, Liquid; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D009132:Muscles / Q000201:enzymology*; D010764:Phosphorylase Kinase / Q000302:isolation & purification* / Q000378:metabolism; D010763:Phosphorylase b / Q000302:isolation & purification* / Q000378:metabolism; D011817:Rabbits
D002413:Cations, Divalent; D007527:Isoenzymes; D010763:Phosphorylase b; D010764:Phosphorylase Kinase; D002118:Calcium