The polysome as a terminal for the creatine phosphate energy shuttle.
The role of the creatine phosphate shuttle in the energetics of muscle protein synthesis in isolated polysomes, from rat hindlimb muscle, was studied. Triton X-100-treated polysomes, following their centrifugation through a 1 M sucrose gradient, contained 38 mU/mg RNA of bound creatine kinase. In the presence of pH 5 enzyme (obtained from rat liver), 0.5 mM ATP, and 1 microM GTP, amino acid (leucine) incorporation by polysomes in the presence of 8 mM creatine phosphate was twice that in the presence of an exogenous ATP regenerating system of 10 mM phospho(enol)pyruvate and 10 U/ml pyruvate kinase. Since added creatine kinase had no effect on incorporation supported by creatine phosphate it is clear that endogenous creatine kinase allows sufficient regeneration of ATP. These data also suggest that nucleoside diphosphokinase must have been associated with the polysome for phosphate was transferred to GTP from [33P]creatine phosphate, and the specific activities of ATP and GTP increased at equal rates, reaching the specific activity of creatine phosphate at 8 min. We conclude that skeletal muscle polysomes have bound creatine kinase activity and they act as terminals for the creatine phosphate energy shuttle. Creatine phosphate regenerates GTP, probably through an intermediate reaction catalyzed by nucleoside diphosphokinase. This provided an added support for the hypothesis of compartmentation of enzymes and substrates and that the transport form of energy between the mitochondria and energy utilizing sites in muscle is creatine phosphate rather than ATP, which extends the general role of the creatine phosphate energy shuttle.
Savabi
F
F
Department of Pharmacology and Nutrition, University of Southern California, School of Medicine, Los Angeles 90033.
Carpenter
C L
CL
Mohan
C
C
Bessman
S P
SP
D000255:Adenosine Triphosphate / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D003402:Creatine Kinase / Q000378:metabolism; D004734:Energy Metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D008297:Male; D009132:Muscles / Q000378:metabolism*; D009701:Nucleoside-Diphosphate Kinase / Q000378:metabolism; D010725:Phosphocreatine / Q000378:metabolism*; D011132:Polyribosomes / Q000378:metabolism*; D014176:Protein Biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains
D000596:Amino Acids; D010725:Phosphocreatine; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D003402:Creatine Kinase; D009701:Nucleoside-Diphosphate Kinase