Deimination of glycogen phosphorylase b by peptidylarginine deiminase. Influence on the kinetical characteristics and dimer-tetramer transition.
The kinetics of the native glycogen phosphorylase b from rabbit skeletal muscle and of the enzyme specifically deiminated by peptidylarginine deiminase have been studied. One arginine residue per phosphorylase b monomer is transformed into citrulline after 3 h of incubation with peptidylarginine deiminase. The maximal velocity of the enzymatic reaction for the modified phosphorylase b is 7-20% higher than that for the native enzyme. Deiminated phosphorylase b, like the native enzyme, shows a positive kinetic cooperativity with respect to glucose-1-phosphate. The affinity of the modified phosphorylase b for the allosteric activator AMP is one order of magnitude higher than that of the native enzyme. Deimination caused a pronounced reduction of the values of [I]0.5 for FMN and glucose, but the sensitivity of the deiminated enzyme to glucose-6-phosphate is much lower than that of the native phosphorylase b. Deiminated phosphorylase b, unlike the native enzyme, shows the positive cooperativity for FMN binding. Deiminated phosphorylase b, unlike the native enzyme, shows less capability to form tetramers in the presence of AMP as compared to the native enzyme.
Eronina
T B
TB
Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia.
Livanova
N B
NB
Chebotareva
N A
NA
Kurganov
B I
BI
Luo
S
S
Graves
D J
DJ
D000249:Adenosine Monophosphate / Q000378:metabolism; D000494:Allosteric Regulation; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001704:Biopolymers; D004789:Enzyme Activation; D005486:Flavin Mononucleotide / Q000378:metabolism; D006867:Hydrolases / Q000378:metabolism*; D007097:Imines / Q000378:metabolism*; D007700:Kinetics; D018482:Muscle, Skeletal / Q000201:enzymology; D010763:Phosphorylase b / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D011817:Rabbits
D000596:Amino Acids; D001704:Biopolymers; D007097:Imines; D000249:Adenosine Monophosphate; D005486:Flavin Mononucleotide; D010763:Phosphorylase b; D006867:Hydrolases; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases