Abstract
Comparison of pig heart aconitase (Kennedy et al., 1972) with yeast (Candida lipolytica) aconitase (Suzuki et al., 1973) reveals similarities in molecular weight and iron content but not in sulphide content. Comparison with the Mildvan & Villafranca (1971) pig heart aconitase preparation reveals differences in iron ligands, specific activity and other properties; these differences possibly arise from protein association as pig heart protein associates under a variety of conditions. The electron spin resonance spectrum, g 4.25, and the low molar relaxivity, 473m(-1).s(-1), of water H(+) suggest the presence of high-spin Fe(III) unco-ordinated to water in the enzyme. The iron chromophore on acid titration at 320nm gives a curve with an inflexion at pH4.2. Ten of 16 expected thiol equivalents are titrated with p-hydroxymercuribenzoate suggesting the presence of cystine as well as cysteine residues. Inhibition of the activation of inactive (activatable) enzyme is sigmoidally related to the molar ratio, p-hydroxymercuribenzoate/enzyme with 10-11mol of mercurial compound causing complete inhibition. Active enzyme, free from activating reagents, requires high molar ratios of mercurial compound for rapid inhibition. In terms of p-hydroxymercuribenzoate the enzyme then lacks an essential thiol group.
MeSH Terms
Aconitate Hydratase/analysis
Amino Acids, Sulfur/analysis
Animals
Candida/enzymology
Cystine/analysis
Electron Spin Resonance Spectroscopy
Enzyme Inhibitors
Hydro-Lyases
Hydrogen-Ion Concentration
Hydroxymercuribenzoates
Iron/analysis
Ligands
Molecular Weight
Myocardium/enzymology
Protein Binding
Sulfhydryl Compounds/analysis
Sulfides/analysis
Swine
Chemicals
Amino Acids, Sulfur
Enzyme Inhibitors
Hydroxymercuribenzoates
Ligands
Sulfhydryl Compounds
Sulfides
Cystine
Iron
Hydro-Lyases
Aconitate Hydratase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gawron O
Kennedy M C
Rauner R A
References (32)
32 references, click to expand
-
Estimation of the molecular weights of proteins by Sephadex gel-filtration.
Biochem J. 1964 May;91(2):222-33
PMID: 4158310
-
On the mechanism of action of aconitase.
J Biol Chem. 1956 May;220(1):193-208
PMID: 13319338
-
alpha-Methyl-cis-aconitic acid. Aconitase substrate. II. Substrate properties and aconitase mechanism.
Biochemistry. 1966 Jul;5(7):2343-50
PMID: 5959457
-
Mechanism of aconitase action. I. The hydrogen transfer reaction.
J Biol Chem. 1967 Apr 25;242(8):1870-9
PMID: 6024777
-
Sub-unit structure and specificity of methionyl-transfer-ribonucleic acid synthetase from Escherichia coli.
Biochem J. 1968 Jun;108(2):281-8
PMID: 4874971
-
Molecular models of metal chelates to illustrate enzymatic reactions.
J Chem Educ. 1968 Aug;45(8):539-40
PMID: 5666356
-
Primary structure of alfalfa ferredoxin.
J Biol Chem. 1969 Feb 10;244(3):981-95
PMID: 5769194
-
Model systems for the iron-sulfur chromophore on nonheme iron proteins.
Biochem Biophys Res Commun. 1969 Jun 6;35(5):634-41
PMID: 5815817
-
The estimation of polypeptide chain molecular weights by gel filtration in 6 M guanidine hydrochloride.
J Biol Chem. 1969 Sep 25;244(18):4989-94
PMID: 5824574
-
Estimation of molecular size and molecular weights of biological compounds by gel filtration.
Methods Biochem Anal. 1970;18:1-53
PMID: 4909316
-
Aspects of enzyme mechanisms studies by nuclear spin relazation induced by paramagnetic probes.
Adv Enzymol Relat Areas Mol Biol. 1970;33:1-70
PMID: 4916855
-
Properties of spinach ferredoxin in anaerobic urea solution: a comparison with the native protein.
Arch Biochem Biophys. 1970 Dec;141(2):456-64
PMID: 4322287
-
The mechanism of aconitase action. I. Preparation, physical properties of the enzyme, and activation by iron (II).
J Biol Chem. 1971 Feb 10;246(3):772-9
PMID: 5542689
-
Succinate dehydrogenase. I. Purification, molecular properties, and substructure.
Biochemistry. 1971 Jun 22;10(13):2509-16
PMID: 4326769
-
The influence of the dimerisation on the stoichiometry of the active center in ferredoxin from Clostridium pasteurianum.
Eur J Biochem. 1971 Sep 13;22(1):57-65
PMID: 5099215
-
An analysis of the electron paramagnetic resonance spectrum of pseudomonas oleovorans rubredoxin. A method for determination of the liganids of ferric iron in completely rhombic sites.
J Biol Chem. 1971 Oct 10;246(19):5877-81
PMID: 4330058
-
Mechanism of aconitase action deduced from crystallographic studies of its substrates.
J Mol Biol. 1968 Dec 14;38(2):149-62
PMID: 5761100
-
1,2,3-DL-tricarboxycyclopentene-1 and other inhibitors of cis-aconitase.
Arch Biochem Biophys. 1971 Dec;147(2):772-80
PMID: 5136110
-
On pig heart aconitase.
Biochem Biophys Res Commun. 1972 May 26;47(4):740-5
PMID: 5026292
-
The mechanism of aconitase action. 3. Detection and properties of enzyme-metal-substrate and enzyme-metal-inhibitor bridge complexes with manganese(II) and iron(II).
J Biol Chem. 1972 Jun 10;247(11):3454-63
PMID: 4337855
-
A novel compound with a planar Fe-S-S-Fe bridge and its possible relation to ferredoxins.
J Am Chem Soc. 1973 Jan 10;95(1):273-4
PMID: 4682896
-
Iron-sulfur proteins: structure and function.
Annu Rev Biochem. 1973;42(0):159-204
PMID: 4599384
-
Iron and aconitase activity.
Biochem J. 1974 Jun;139(3):709-14
PMID: 4852570
-
Sulphydryl-disulphide relationships in the induction of gels in proteins by urea.
Nature. 1951 Apr 14;167(4250):592-3
PMID: 14826861
-
The purification of aconitase.
Biochem J. 1954 Jan;56(1):99-105
PMID: 13126098
-
Determination of--SH groups in proteins.
Methods Biochem Anal. 1962;10:43-70
PMID: 13967409
-
THE ABSOLUTE STEREOCHEMICAL COURSE OF CITRIC ACID BIOSYNTHESIS.
Proc Natl Acad Sci U S A. 1963 Nov;50:981-8
PMID: 14082366
-
THE SPECIFIC BINDING OF IRON(III) AND COPPER(II) TO TRANSFERRIN AND CONALBUMIN.
Biochim Biophys Acta. 1963 Sep 24;75:203-22
PMID: 14083902
-
STUDIES ON THE CHEMICAL NATURE OF CLOSTRIDIAL FERREDOXIN.
J Biol Chem. 1963 Dec;238:3899-913
PMID: 14086723
-
THE DISSOCIATION AND ASSOCIATION OF PROTEIN STRUCTURES.
Adv Protein Chem. 1963;18:123-226
PMID: 14151995
-
EQUILIBRIUM ULTRACENTRIFUGATION OF DILUTE SOLUTIONS.
Biochemistry. 1964 Mar;3:297-317
PMID: 14155091
-
The gel-filtration behaviour of proteins related to their molecular weights over a wide range.
Biochem J. 1965 Sep;96(3):595-606
PMID: 5862401