Glycogen phosphorylase b and phosphorylase kinase binding to glycogen under molecular crowding conditions. Inhibitory effect of FAD

• Published in: Biochemistry (Moscow) Vol. 74; no. 5; pp. 562 - 568
• Main Authors: Chebotareva, N. A., Meremyanin, A. V., Makeeva, V. F., Eronina, T. B., Kurganov, B. I.
• Format: Journal Article
• Language: English
• Published: Dordrecht SP MAIK Nauka/Interperiodica 01.05.2009; Springer Nature B.V
• Subjects: Animals; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; Enzyme Inhibitors - metabolism; Enzymes; Flavin-Adenine Dinucleotide - metabolism; Glycogen - metabolism; Glycogen Phosphorylase, Muscle Form - chemistry; Glycogen Phosphorylase, Muscle Form - metabolism; Life Sciences; Macromolecular Substances - chemistry; Macromolecular Substances - metabolism; Microbiology; Molecular biology; Particle Size; Phosphorylase Kinase - chemistry; Phosphorylase Kinase - metabolism; Protein Binding; Rabbits; dynamic light scattering; crowding; FAD; phosphorylase kinase; glycogen; trimethylamine N-oxide; glycogen phosphorylase
• ISSN: 0006-2979; 1608-3040
• DOI: 10.1134/S0006297909050125

Dynamic light scattering was used to study the interaction of phosphorylase kinase (PhK) and glycogen phos-phorylase b (Ph b ) from rabbit skeletal muscle with glycogen under molecular crowding conditions arising from the presence of 1 M trimethylamine N-oxide and at physiological ionic strength. The mean value of hydrodynamic radius of the initial glycogen particles was 52 nm. Crowding stimulated Ph b and PhK combined binding on glycogen particles. Two-stage character of PhK binding to glycogen particles containing adsorbed Ph b was found in the presence of the crowding agent. At the initial stage, limited size particles with hydrodynamic radius of ∼220 nm are formed, whereas the second stage is accompanied by linear growth of hydrodynamic radius. Flavin adenine dinucleotide (FAD) selectively inhibited PhK binding at the second stage. The data indicate that in the first stage Ph b is involved in PhK binding by glycogen particles containing adsorbed Ph b , whereas PhK binding in the second stage does not involve Ph b .