Reactivity of the Active-centre Lysine Residue of Rabbit Muscle Aldolase

Overview

Journal Biochem J

Specialty Biochemistry

Date 1974 Feb 1

PMID 4856792

Citations 1

Authors

P J Anderson

H Kaplan

Affiliations

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Abstract

The method of competitive labelling with [(3)H]acetic anhydride as the labelling reagent was used to determine the properties of the active-centre lysine residue of rabbit muscle aldolase. This residue is much less reactive than a normal exposed lysine residue towards this reagent, and its reactive properties did not parallel the pH-activity profile for aldolase. At higher pH values it became reactive, but this was shown to be due to disruption of the enzyme structure. The binding of the competitive inhibitor phosphate did not alter the reactive properties. It is concluded that the active-centre lysine has an apparent pK(a) greater than 11.5 and probably is made nucleophilic during the catalytic process, perhaps by proton abstraction.

Citing Articles

Intramolecular ionic interactions of lysine residues and a possible folding domain in fructose diphosphate aldolase.

LAMBERT J, Perham R, Coggins J Biochem J. 1977; 161(1):63-71.

PMID: 851425 PMC: 1164474. DOI: 10.1042/bj1610063.

References

Cash D, Wilson I . The cyanide adduct of the aldolase dihydroxyacetone phosphate imine. J Biol Chem. 1966; 241(18):4290-2. View

Gibbons I, Anderson P, Perham R . Amino acid sequence homology in the active site of rabbit and sturgeon muscle aldolases. FEBS Lett. 1970; 10(1):49-53. DOI: 10.1016/0014-5793(70)80413-6. View

Blow D, Birktoft J, Hartley B . Role of a buried acid group in the mechanism of action of chymotrypsin. Nature. 1969; 221(5178):337-40. DOI: 10.1038/221337a0. View

MEHLER A, Cusic Jr M . Aldolase reaction with sugar diphosphates. Science. 1967; 155(3766):1101-3. DOI: 10.1126/science.155.3766.1101. View

Blow D, Steitz T . X-ray diffraction studies of enzymes. Annu Rev Biochem. 1970; 39:63-100. DOI: 10.1146/annurev.bi.39.070170.000431. View

Grazi E, Rowley P, Cheng T, TCHOLA O, HORECKER B . The mechanism of action of aldolases. III. Schiff base formation with lysine. Biochem Biophys Res Commun. 1962; 9:38-43. DOI: 10.1016/0006-291x(62)90083-9. View

DOUNCE A, BARNETT S, BEYER G . Further studies on the kinetics and determination of aldolase. J Biol Chem. 1950; 185(2):769-80. View

Kaplan H . Determination of the ionization constants and reactivities of the amino-termini of -chymotrypsin. J Mol Biol. 1972; 72(1):153-62. DOI: 10.1016/0022-2836(72)90076-9. View

Brown J, Hartley B . Location of disulphide bridges by diagonal paper electrophoresis. The disulphide bridges of bovine chymotrypsinogen A. Biochem J. 1966; 101(1):214-28. PMC: 1270086. DOI: 10.1042/bj1010214. View

10.

Lai C, Oshima T . Studies on the structure of rabbit muscle aldolase. 3. Primary structure of the BrCN peptide containing the active site. Arch Biochem Biophys. 1971; 144(1):363-74. DOI: 10.1016/0003-9861(71)90489-9. View

11.

Schmidt Jr D, WESTHEIMER F . PK of the lysine amino group at the active site of acetoacetate decarboxylase. Biochemistry. 1971; 10(7):1249-53. DOI: 10.1021/bi00783a023. View

12.

Rutter W . EVOLUTION OF ALDOLASE. Fed Proc. 1964; 23:1248-57. View

13.

Kaplan H, Stevenson K, Hartley B . Competitive labelling, a method for determining the reactivity of individual groups in proteins. The amino groups of porcine elastase. Biochem J. 1971; 124(2):289-99. PMC: 1177144. DOI: 10.1042/bj1240289. View

14.

Anderson P, Gibbons I, Perham R . A comparative study of the structure of muscle fructose 1,6-diphosphate aldolases. Eur J Biochem. 1969; 11(3):503-9. DOI: 10.1111/j.1432-1033.1969.tb00802.x. View

15.

Model P, PONTICORVO L, Rittenberg D . Catalysis of an oxygen-exchange reaction of fructose 1,6-diphosphate and fructose 1-phosphate with water by rabbit muscle aldolase. Biochemistry. 1968; 7(4):1339-47. DOI: 10.1021/bi00844a014. View