[Reconstruction of muscle glycogen phosphorylase b from an apoenzyme and pyridoxal-5'-phosphate and its analogs. Interaction of apophosphorylase and the reconstructed enzyme with specific ligands].
Sedimentation methods were used to study the effects of modification of the pyridoxal-5'-phosphate (PLP) molecule at the 5th position on the affinity of reconstituted muscle glycogen phosphorylase b for the substrate (glycogen) and the allosteric inhibitor (FMN) as well as on the enzyme capacity to association induced by AMP. Reconstituted phosphorylase b was obtained with PLP analogs containing at the 5th position -CH2-CH2-COOH (analog I), trans-CH=CH-COOH (analog II) or -C identical to COOH (analog III) residues. Reconstitution of phosphorylase b is accompanied by the recovery of the enzyme quaternary structure. Phosphorylase b reconstituted with PLP or analogs I, II and III is not distinguished practically from the native enzyme in its affinity for glycogen. Substitution of the native coenzyme in the phosphorylase molecule with any tested PLP analog leads to lower enzyme affinity for FMN. Microscopic dissociation constants of the FMN-enzyme complexes increase in the following order: enzyme.I < enzyme.II < enzyme.III. Phosphorylase b reconstituted with analogs I, II and III differs substantially from the native enzyme in its capacity to association in the presence of 1 mM AMP: the reconstituted enzyme is represented practically by only the tetrameric form.
Chebotareva
N A
NA
Sugrobova
N P
NP
Bulanova
L N
LN
Poznanskaia
A A
AA
Kurganov
B I
BI
Gunar
V I
VI
D000249:Adenosine Monophosphate / Q000494:pharmacology; D000494:Allosteric Regulation; D000818:Animals; D005486:Flavin Mononucleotide / Q000494:pharmacology; D006003:Glycogen / Q000378:metabolism; D008024:Ligands; D018482:Muscle, Skeletal / Q000201:enzymology*; D010763:Phosphorylase b / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000378:metabolism*; D011817:Rabbits; D013379:Substrate Specificity
D008024:Ligands; D000249:Adenosine Monophosphate; D011732:Pyridoxal Phosphate; D005486:Flavin Mononucleotide; D006003:Glycogen; D010763:Phosphorylase b