Disulfide bond formation between the active-site thiol and one of the several free thiol groups of chymopapain

Ibolya Kóródi, Bence Asbóth, L. Polgár

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Abstract

Chymopapain, a cysteine protease of papaya latex, has been purified with the use of fast protein liquid chromatography. Two homogeneous fractions were analyzed for thiol content and thiol reactivity. It was found that peak 1 and peak 2 contained two and three thiol groups, respectively, per mole of enzyme. This result is inconsistent with the general belief that chymopapain contains one essential and one nonessential thiol group and suggests that a significant portion of the thiol groups was oxidized in the previous preparations. Such an oxidation can account for some of the inconsistent results reported in the literature. An irreversibly oxidized nonessential thiol group may modify the catalytic function of chymopapain especially if it is close to the active site. That one thiol group resides indeed in the vicinity of the essential thiol group is clearly demonstrated by the biphasic reactions of chymopapain with disulfide compounds such as 2,2′-dipyridyl disulfide and 5,5′-dithiobis(2-nitrobenzoate). In the first step of these reactions a mixed disulfide is formed between the enzyme and the reactant, which is followed by a first-order, intramolecular reaction leading to the liberation of the second half of the disulfide compound. Furthermore, on addition of one Hg2+ ion, 2 mol of thiol group, one essential and one nonessential, disappears concomitantly. Formation of a disulfide bond between the catalytically competent thiol group and another free thiol group of chymopapain under physiological conditions may be of regulatory importance.

Original languageEnglish
Pages (from-to)6895-6900
Number of pages6
JournalBiochemistry
Volume25
Issue number22
Publication statusPublished - 1986

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Chymopapain
Sulfhydryl Compounds
Disulfides
Catalytic Domain
Carica
Cysteine Proteases
Liquid chromatography
Latex
Enzymes
Liquid Chromatography

ASJC Scopus subject areas

  • Biochemistry

Cite this

Disulfide bond formation between the active-site thiol and one of the several free thiol groups of chymopapain. / Kóródi, Ibolya; Asbóth, Bence; Polgár, L.

In: Biochemistry, Vol. 25, No. 22, 1986, p. 6895-6900.

Research output: Contribution to journalArticle

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