Comparative study of the interactions AMP-phosphorylase b and AMP analogues-phosphorylase b.
The different effects induced by AMP and its analogues on the tertiary structure and the coenzyme environment of phosphorylase b were studied by titration of the slowly reacting thiol groups and by quenching of the coenzyme fluorescence, respectively, to determine the possible differences that activate phosphorylase b. The following results were obtained: The coenzyme environment depends upon the nucleotide studied. AMP, when bound to its first site, opens the coenzyme pocket. The slow cysteines were shielded by the nucleotides against their DTNB titration depending on the nucleotide studied. The enzyme difference spectra in presence of the nucleotide showed that the negative band of 260 nm is similar for all nucleotides possessing the same base, but the positive band obtained in the presence of AMP was diminished when other nucleotides were present.
D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D003067:Coenzymes / Q000378:metabolism; D004789:Enzyme Activation; D066298:In Vitro Techniques; D010763:Phosphorylase b / Q000378:metabolism*; D006005:Phosphorylases / Q000378:metabolism*; D011487:Protein Conformation; D011817:Rabbits; D013056:Spectrophotometry, Ultraviolet; D013438:Sulfhydryl Compounds
D003067:Coenzymes; D013438:Sulfhydryl Compounds; D000249:Adenosine Monophosphate; D010763:Phosphorylase b; D006005:Phosphorylases