Functional arginyl residues in carboxypeptidase A. Modification with butanedione

Riordan, James

doi:10.1021/bi00744a020

Cited by 368 publications

(237 citation statements)

References 23 publications

Supporting

Mentioning

204

Contrasting

Unclassified

Order By: Relevance

“…Thermolysin possesses multiple binding sites for the leaving, group, S], S~...S~, located in an extended cleft of the enzyme while carboxypeptidase A possesses only a single binding site for the leaving group i.e. S], which is located in a dead-end pocket in the enzyme (14,16,17). The active sites of the serine proteases are similarly located in clefts (15,18), and in analogy with carboxypeptidase A we might expect dead-end structures in the serine carboxypeptidases.…”

Section: Discussionmentioning

confidence: 99%

Influence of guanidine derivatives on the specificity of malt carboxypeptidase

Breddam

Ottesen

1983

Carlsberg Res. Commun.

View full text Add to dashboard Cite

Malt carboxypeptidase (Carlsberg Res. Commun. 48, 217-230, 1983) catalyzes hydrolysis of ester substrates with the general formula Bz-X-OMe with preference for substrates where X = Phe, Arg or Lys over those where the X-position is occupied by an amino acid residue with a neutral or acid aliphatic side chain. The rapid hydrolysis of Bz-Phe-OMe is mainly due to a high k~, value while for Bz-Arg-OMe it is mainly due to a low Km value. Addition of salts result in decreased rates of hydrolysis of substrates where X = Lys or Arg (positively charged) and increased rates of hydrolysis of substrates where X = Phe or Leu (hydrophobic).Guanido compounds influence the enzymatic properties of malt carboxypeptidase. Kinetic data for the hydrolysis of various substrates indicate that phenylguanidine binds to the St' binding site of the enzyme in such a manner that the rate of cleavage of ester or amide substrates with large groups in the P( position, i.e. -OEt, -OPt, -OBu, -Ala-OH, -Gly-NH2, is reduced while substrates with small groups in this position i.e. -OMe, -NH2 are hydrolyzed with increased rates. These observations have applications in deamidation reactions since the presence of phenylguanidine in the reaction medium results in a significantly increased yield of the deamidated product.Abbreviations: Ac = acetyl; BiGu = 2-guanidobenzimidazol; Bu = butyl; Bz = benzoyl; DFP = diisopropylphosphorofluoridate; EDTA = ethylene diamine tetraacetic acid sodium salt; FA = furylacryloyl; Gu = guanidine hydrochloride; Hepes = N-2-hydroxyethylpiperazine-N'-2-ethane sulfonic acid; HPLC = high performance liquid chromatography; MeGu = methyl guanidine hydrochloride; Mes = 2-(N-morpholino)ethane sulfonic acid; PhGu = phenylguanidine hydrogencarbonate; PpGu = l-(3-phenylpropylamino) guanidine hydrochloride; Pr = propyl; TEAP = triethyl ammonium phosphate; "Iris = tris(hydroxy methyl) aminomethane; Z = carbobenzoxy. Abbreviations of amino acids, amino acid derivatives and peptides are according to the guidelines of the IUPAC-IUB Commision on Biochemical Nomenclature. The binding notation for the enzyme and the substrates is that of SCHECHTER and BERGER. Accordingly, amino acid residues in the substrate are referred to as P,,P2....P~ in the aminoterminal direction away from the scissible bond and P~ for the carboxy-terminal residue that is hydrolyzed off.

show abstract

Section: Discussionmentioning

confidence: 99%

Influence of guanidine derivatives on the specificity of malt carboxypeptidase

Breddam

Ottesen

1983

Carlsberg Res. Commun.

View full text Add to dashboard Cite

show abstract

“…Along this line, selective chemical modifications of strategic amino acids in the ADP/ATP carrier have already provided interesting data [4,5,8]. The penetrant reagents used so tar and their main targets include NEM (SH groups) [91, HNB (tryptophanyl residues) [10], phenylglyoxal and butanedione (arginyl residues [ 11,12]). Inactivation of the binding properties of the ADP/ATP carrier by UV light was possibly due to modification of tryptophanyl residues [4].…”

Section: Introductionmentioning

confidence: 99%

Atractyloside and bongkrekic acid sites in the mitochondrial ADP/ATP carrier protein

1981

View full text Add to dashboard Cite

“…The results described subsequently focus on the first phase of the reaction. It should be pointed out that the reactions of glutamate dehydrogenase with 2,3-butanedione reported in this paper were all conducted in 0.02 M potassium phosphate buffer, despite the report of Riordan [8] that borate buffer enhances the reaction rate of 2,3-butanedione with arginine. Johnson and Smith [22a] have shown that borate can form complexes with cis-diols such as those found in the ribose moieties of NAD and ADP; therefore borate was not used in the reaction mixtures of glutamate dehydrogenase and 2,3-butanedione in order to avoid complicating the interpretation of any effect of added nucleotides on the course of the reaction.…”

Section: Inactivation Of Glutamate Dehydrogenase By 23-butanedionementioning

confidence: 68%

The Importance of Arginine Residues in the Catalytic and Regulatory Functions of Bovine‐Liver Glutamate Dehydrogenase

Pal

Colman

1976

European Journal of Biochemistry

View full text Add to dashboard Cite

Bovine liver glutamate dehydrogenase reacts rapidly with 2,3-butanedione to yield modified enzyme with 29% of its original maximum activity, but no change in its Michaelis constants for substrates and coenzymes. No significant reduction in the inactivation rate is produced by the addition of the allosteric activator ADP or inhibitor GTP, while partial protection against inactivation is provided by the coenzyme NAD' or substrate 2-oxoglutarate when added separately. The most marked decrease in the rate of inactivation (about 10-fold) is provided by the combined addition of NAD+ and 2-oxoglutarate, suggesting that modification takes place in the region of the active site. Reaction with 2,3-butanedione also results in loss of the ability of the enzyme to be activated by ADP. Addition of ADP (but not NAD', 2-oxoglutarate or GTP) to the incubation mixture protects markedly against the loss of activatability by ADP. It is concluded that 2,3-butanedione produces two distinguishable effects on glutamate dehydrogenase : a relatively specific modification of the regulatory ADP site and a distinct modification in the active center. Reaction of two arginyl residues per peptide chain appears to be responsible for disruption of the ADP activation property of the enzyme, while alteration of a maximum of five arginyl residues can be related to the reduction of maximum catalytic activity. Electrostatic interactions between the positively charged arginine groups and the negatively charged substrate, coenzyme and allosteric purine nucleotide may be important for the normal function of glutamate dehydrogenase.Arginine residues have been shown to participate in the binding sites for coenzyme [l -71 or substrate [8 -1 I] of a variety of enzymes. The common feature of the enzymes for which arginine is essential seems to be the catalysis of a reaction involving a negatively charged substrate or coenzyme. Glutamate dehydrogenase [L-glutamate : NAD(P) oxidoreductase (deaminating)] not only has negatively charged substrates and coenzymes, but is also regulated by negatively charged purine nucleotides which bind at distinct allosteric sites. Thus, it has several regions which might be postulated to include arginine. Chemical modification studies have implicated tyrosyl, lysyl and methionyl residues in the function of glutamate dehydrogenase [12-191, but little attention has been focused on the role of arginyl residues. While this work was in progress, Blumenthal and Smith reported, on the basis of reaction with 3,2 cyclohexanedione, that arginyl residues play important Enzyme. Bovine liver glutamate dehydrogenase or gluta am ate : NAD(P) oxidoreductase (deaniinating) (EC 1.4.1.3). roles in the binding of NADP or NAD by the glutamate dehydrogenases of Neurospora and bovine liver [20]. The present paper, utilizing chemical modification of bovine liver glutamate dehydrogenase by 2,3-butanedione, reveals that arginyl residues are involved both in the regulation by purine nucleotides and in the catalytic function of the enzyme. EXPERIMENTAL PR...

show abstract

Functional arginyl residues in carboxypeptidase A. Modification with butanedione

Cited by 368 publications

References 23 publications

Influence of guanidine derivatives on the specificity of malt carboxypeptidase

Influence of guanidine derivatives on the specificity of malt carboxypeptidase

Atractyloside and bongkrekic acid sites in the mitochondrial ADP/ATP carrier protein

The Importance of Arginine Residues in the Catalytic and Regulatory Functions of Bovine‐Liver Glutamate Dehydrogenase

Contact Info

Product

Resources

About