Differential effects of cysteine on protein and coenzyme A synthesis in rat heart.
The effect of cysteine availability on protein and coenzyme A (CoA) synthesis in perfused rat heart was incompletely evaluated in earlier experiments because rapid conversion of cysteine to cystine occurred when the perfusion buffer was oxygenated. This conversion was minimized by addition of an excess of reducing agents such as dithiothreitol or mercaptodextran or by provision of bathocuproine disulfonate, a copper chelator. Dithiothreitol was not a suitable protective agent because it reduced ATP and creatine phosphate contents. Perfusion of hearts with [35S]cystine or [35S]cysteine in the presence of mercaptodextran resulted in a 22-fold or 5-fold increase, respectively, in incorporation of [35S] into protein and a 5-fold or 8-fold increase, respectively, in incorporation into CoA compared with hearts supplied [35S]cystine or [35S]cysteine without the reducing agent. When compared with hearts perfused at an aortic pressure of 90 mmHg with bicarbonate buffer that contained 15 mM glucose, 25 mU insulin/ml, 0.4 mM [14C]phenylalanine, no cysteine and plasma levels of other amino acids, provision of 0.09 or 0.2 mM cysteine alone or in the presence of mercaptodextran, or bathocuproine disulfonate enhanced rates of protein synthesis 16-35%. When 0.2 mM cysteine was added to bicarbonate buffer containing 7 microM pantothenic acid, supplementation with mercaptodextran or bathocuproine disulfonate was required to raise CoA content. These results indicated that an exogenous supply of cysteine was needed to maintain maximal rates of protein and CoA synthesis in the perfused rat heart. Protective compounds were required to obtain the cysteine effect on CoA but not on protein synthesis.
D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology; D002021:Buffers; D002250:Carbon Radioisotopes; D003065:Coenzyme A / Q000096:biosynthesis*; D003545:Cysteine / Q000494:pharmacology*; D003553:Cystine; D003911:Dextrans / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D006321:Heart / Q000187:drug effects*; D008297:Male; D009206:Myocardium / Q000378:metabolism; D010205:Pantothenic Acid; D010477:Perfusion; D010618:Phenanthrolines / Q000494:pharmacology; D010649:Phenylalanine; D014176:Protein Biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013462:Sulfur Radioisotopes
D001639:Bicarbonates; D002021:Buffers; D002250:Carbon Radioisotopes; D003911:Dextrans; D010618:Phenanthrolines; D013438:Sulfhydryl Compounds; D013462:Sulfur Radioisotopes; D010205:Pantothenic Acid; C004232:mercaptodextran; D010649:Phenylalanine; D003553:Cystine; C028559:bathocuproine sulfonate; D003545:Cysteine; D003065:Coenzyme A; D004229:Dithiothreitol