Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:4.1.2.13 (aldolase)
 3,461 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Chemical modifications of Class I aldolases from Trypanosoma brucei, rabbit muscle and Staphylococcus aureus with carboxypeptidase A, glyceraldehyde 3-phosphate and cysteine-specific reagents revealed the following differences between the three homologous enzymes. Aldolase from S. aureus was not affected by any of these reagents. Carboxypeptidase-A treatment of rabbit-muscle and T. brucei aldolase inhibited the activity of both enzymes towards fructose-1,6-bisphosphate (Fru(1,6)P2), while the activity towards fructose-1-phosphate (Fru-1-P) was affected only in the case of the trypanosomal enzyme. Moreover carboxypeptidase-A treatment reduced the turnover numbers of these two aldolases for both Fru(1,6)P2 and Fru-1-P to a similar level. Glyceraldehyde 3-phosphate, in the absence of dihydroxyacetone phosphate, also inactivated aldolases from rabbit muscle and T. brucei with second order rate constants of 1054 and 254 min-1 M-1, respectively. Using 5,5'-dithiobis-(2-nitrobenzoic acid) with rabbit-muscle aldolase, a total of 4 thiol groups could be titrated per subunit, resulting in a total inactivation. The presence of substrate completely protected the enzyme from inactivation. Methyl methanethiosulfonate also reacted with four cysteine residues, but this led to very little inactivation. This indicates that the inactivation by modification with DTNB is due to conformational changes in the enzyme. In T. brucei aldolase only one thiol group could be titrated with methyl methanesulfonate and there was no loss of activity. With 5,5'-dithiobis-(2-nitrobenzoic acid) five cysteines were titrated with an immediate and complete loss of activity.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Chemical modification of fructose bisphosphate aldolase from Trypanosoma brucei compared to aldolase from rabbit muscle and Staphylococcus aureus. 185 80

At pH 7.0 and 25 degrees C, NBF-Cl (4-chloro-7-nitrobenzofurazan) reacts rapidly with rabbit muscle aldolase (D-fructose-1,6-bisphosphate D-glyceraldehyde-3-phosphate-lyase, EC 4.1.2.13) to yield a product with an absorption maximum at 402 nm, which is shifted to 422 nm upon acid denaturation. The reaction involves arylation of a single cysteine residue per subunit of tetrameric aldolase, as shown by the molar absorptivity of NBF-aldolase and by titration of sulfhydryl groups of the enzyme with 5,5'-dithiobis(2-nitrobenzoic acid), DTNB. The site of arylation appears to be Cys-237, which is the cysteine residue that reacts most rapidly with DTNB. The site of arylation appears to be Cys-237, which is the cysteine residue that reacts most rapidly with DTNB, and which is not essential for aldolase activity. Arylation of the enzyme is 13-20-times more rapid than that of model compounds. The relatively high rate of arylation is not due to medium effects, to an anomalously low pKa of Cys-237, or to the presence of a binding site for NBF-Cl, and is tentatively assigned to acid-base catalysis by other functional groups in the vicinity of the reactive sulfhydryl group. The NBF-Cl reaction provides the most efficient means of titrating Cys-237 residues in rabbit muscle aldolase.
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PMID:Selective arylation of cysteine-237 of rabbit muscle aldolase with 4-chloro-7-nitrobenzofurazan. 705 73