[Purification, quaternary structure and regulatory properties of phosphorylase kinase from pigeon skeletal muscle].
Using DEAE-Toyopearl column chromatography, a preparation of pigeon skeletal muscle phosphorylase kinase was obtained in a state approaching homogeneity. The molecular mass of the native enzyme (1320 kDa) and the subunit formula (alpha beta gamma delta)4 are similar to those of rabbit and chicken counterparts. Both red and white pigeon skeletal muscle isozymes contain the alpha'-subunit instead of alpha. Gradient SDS-PAGE electrophoresis revealed small but well-reproducible differences in the molecular masses of rabbit, chicken and pigeon muscle beta- and gamma-subunits. The activity ratio at pH 6.8/8.2 is 0.06-0.15 for different preparations of phosphorylase kinase b. The activity of pigeon muscle phosphorylase kinase b is Ca2+-dependent. The [Ca2+]0.5 value at pH 7.0 is 20 microM, which exceeds that for the chicken muscle enzyme by two orders of magnitude. In the presence of Ca2+, pigeon phosphorylase kinase b is activated 4-fold by saturating concentrations of calmodulin and troponin C. Pigeon muscle phosphorylase b is activated 3-5-fold during autophosphorylation or phosphorylation by the catalytic subunit of cAMP-dependent protein kinase.
Morozov
V E
VE
Eronina
T B
TB
Andreeva
I E
IE
Silonova
G V
GV
Solov'eva
N V
NV
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000502:physiology; D002645:Chickens; D010856:Columbidae / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D046911:Macromolecular Substances; D009132:Muscles / Q000201:enzymology*; D010764:Phosphorylase Kinase / Q000032:analysis / Q000302:isolation & purification*; D010766:Phosphorylation; D011817:Rabbits; D014336:Troponin / Q000502:physiology; D019209:Troponin C
D002147:Calmodulin; D046911:Macromolecular Substances; D014336:Troponin; D019209:Troponin C; D010764:Phosphorylase Kinase; D002118:Calcium
Ochistka, chetvertichnaia struktura i reguliatornye svoĭstva kinazy fosforilazy iz skeletnykh myshts golubia.