Isoelectrofocusing of rat muscle adenylosuccinase.
Adenylosuccinase catalyses the conversion of adenylosuccinic acid to AMP and fumarate. We have developed a coupled enzyme staining procedure applicable to nitrocellulose blots after agarose gel isoelectrofocusing of rat muscle adenylosuccinase. The coupling enzymes, fumarase (fumarate to L-malate) and malic enzyme (L-malate to pyruvate and NADPH), are adsorbed to nitrocellulose prior to blotting. The NADPH, mediated by phenazine methosulfate, converts a tetrazolium salt to its blue formazan. This procedure demonstrated that rat muscle adenylosuccinase consists of three isomeric forms present in similar amounts.
Mack
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H. L. Snyder Memorial Research Foundation, Winfield, KS.
Smith
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D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000264:Adenylosuccinate Lyase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D005260:Female; D005649:Fumarate Hydratase / Q000378:metabolism; D007525:Isoelectric Focusing / Q000379:methods*; D007527:Isoenzymes; D008773:Methylphenazonium Methosulfate / Q000378:metabolism; D009132:Muscles / Q000201:enzymology*; D009249:NADP / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling
D007527:Isoenzymes; C012168:adenylosuccinate; D008773:Methylphenazonium Methosulfate; D000249:Adenosine Monophosphate; D009249:NADP; D005649:Fumarate Hydratase; D000264:Adenylosuccinate Lyase