Identification of the C-1-phosphate-binding arginine residue of rabbit-muscle aldolase. Isolation of 1,2-cyclohexanedione-labeled peptide by chemisorption chromatography.
The arginine-specific reagent 1,2-cyclohexanedione reacts selectively with the arginine residue of the C-1-phosphate-binding site of aldolase and inactivates the enzyme. The labeled peptide isolated from tryptic digests of inactivated aldolase was found to correspond to the sequence Leu-43 to Arg-56, the residue modified by cyclohexanedione being Arg-55. This peptide was absent form digests of aldolase treated in the same way but protected from inactivation by the presence of substrate, thus correlating modification of Arg-55 with loss of activity. Selective isolation ofthe peptide containing the modified arginine residue was effected by chemisorption chromatography on boric acid gel, a procedure exploiting the specific interaction of matrix-bound boric acid groups with vicinal cis-hxdroxyl groups of cyclohexanedione-modified arginine side chains.
D000818:Animals; D001120:Arginine / Q000032:analysis*; D001665:Binding Sites; D002850:Chromatography, Gel; D003510:Cyclohexanes; D003512:Cyclohexanones; D005634:Fructose-Bisphosphate Aldolase; D009132:Muscles / Q000201:enzymology*; D009943:Organophosphorus Compounds / Q000378:metabolism*
D003510:Cyclohexanes; D003512:Cyclohexanones; D009943:Organophosphorus Compounds; C018031:1,2-cyclohexanedione; D001120:Arginine; D005634:Fructose-Bisphosphate Aldolase