Autoxidizability of beef heart cytochrome c1 lacking the hinge protein c1-c

Kuniaki Mukai, Hiroshi Matsubara

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The autoxidizability of beef heart cytochrome c1 was investigated in terms of the integrity of the binding of the hinge protein to the heme subunit. Cytochrome c1 was isolated as a subcomplex consisting of the heme subunit and the hinge protein. Treatment of the cytochrome c1 subcomplex with p-chloromercuribenzoate (pCMB) under mild conditions lessened the binding strength between the two subunits. They were dissociated on polyacrylamide gel electrophoresis (PAGE) under non-denaturing conditions, but were not separated by gel filtration chromatography. The pCMB-treated subcomplex had a slight autoxidizabiity. This was repressed to the level of the native subeomplex, when the mercurial compound bound to the subcomplex was removed by the addition of 2-mercaptoethanol. Concomitantly, the less stable binding between the subunits was apparently reversed to the native state. After pCMB treatment of the subcomplex, the heme subunit recovered from PAGE showed marked autoxidizabiity, even if it was treated with 2-mercapto-ethanol. Addition of cholate repressed the autoxidizability of the heme subunit after the removal of the mercurial compound. These results confirmed that the stable binding of the hinge protein to the heme subunit was essential for the non-autoxidizability of cytochrome c1 subcomplex. In addition, it was suggested that cysteinyl residues in the subcomplex must be involved to a great extent in the stable binding between the two subunits.

Original languageEnglish
Pages (from-to)1165-1173
Number of pages9
JournalJournal of Biochemistry
Volume100
Issue number5
Publication statusPublished - 1986 Oct
Externally publishedYes

Fingerprint

Cytochromes c1
Beef
Hinges
Heme
Chloromercuribenzoates
Proteins
Protein
Mercaptoethanol
Electrophoresis
Gels
Carrier Proteins
Polyacrylates
Cholates
Native Polyacrylamide Gel Electrophoresis
Protein Subunits
Ethanol
Chromatography
Integrity
Filtration
Gel Chromatography

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Physiology (medical)
  • Radiology Nuclear Medicine and imaging
  • Molecular Biology
  • Biochemistry

Cite this

Autoxidizability of beef heart cytochrome c1 lacking the hinge protein c1-c. / Mukai, Kuniaki; Matsubara, Hiroshi.

In: Journal of Biochemistry, Vol. 100, No. 5, 10.1986, p. 1165-1173.

Research output: Contribution to journalArticle

@article{f01e9f0d1311487faeb48d2660876a63,
title = "Autoxidizability of beef heart cytochrome c1 lacking the hinge protein c1-c",
abstract = "The autoxidizability of beef heart cytochrome c1 was investigated in terms of the integrity of the binding of the hinge protein to the heme subunit. Cytochrome c1 was isolated as a subcomplex consisting of the heme subunit and the hinge protein. Treatment of the cytochrome c1 subcomplex with p-chloromercuribenzoate (pCMB) under mild conditions lessened the binding strength between the two subunits. They were dissociated on polyacrylamide gel electrophoresis (PAGE) under non-denaturing conditions, but were not separated by gel filtration chromatography. The pCMB-treated subcomplex had a slight autoxidizabiity. This was repressed to the level of the native subeomplex, when the mercurial compound bound to the subcomplex was removed by the addition of 2-mercaptoethanol. Concomitantly, the less stable binding between the subunits was apparently reversed to the native state. After pCMB treatment of the subcomplex, the heme subunit recovered from PAGE showed marked autoxidizabiity, even if it was treated with 2-mercapto-ethanol. Addition of cholate repressed the autoxidizability of the heme subunit after the removal of the mercurial compound. These results confirmed that the stable binding of the hinge protein to the heme subunit was essential for the non-autoxidizability of cytochrome c1 subcomplex. In addition, it was suggested that cysteinyl residues in the subcomplex must be involved to a great extent in the stable binding between the two subunits.",
author = "Kuniaki Mukai and Hiroshi Matsubara",
year = "1986",
month = "10",
language = "English",
volume = "100",
pages = "1165--1173",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "5",

}

TY - JOUR

T1 - Autoxidizability of beef heart cytochrome c1 lacking the hinge protein c1-c

AU - Mukai, Kuniaki

AU - Matsubara, Hiroshi

PY - 1986/10

Y1 - 1986/10

N2 - The autoxidizability of beef heart cytochrome c1 was investigated in terms of the integrity of the binding of the hinge protein to the heme subunit. Cytochrome c1 was isolated as a subcomplex consisting of the heme subunit and the hinge protein. Treatment of the cytochrome c1 subcomplex with p-chloromercuribenzoate (pCMB) under mild conditions lessened the binding strength between the two subunits. They were dissociated on polyacrylamide gel electrophoresis (PAGE) under non-denaturing conditions, but were not separated by gel filtration chromatography. The pCMB-treated subcomplex had a slight autoxidizabiity. This was repressed to the level of the native subeomplex, when the mercurial compound bound to the subcomplex was removed by the addition of 2-mercaptoethanol. Concomitantly, the less stable binding between the subunits was apparently reversed to the native state. After pCMB treatment of the subcomplex, the heme subunit recovered from PAGE showed marked autoxidizabiity, even if it was treated with 2-mercapto-ethanol. Addition of cholate repressed the autoxidizability of the heme subunit after the removal of the mercurial compound. These results confirmed that the stable binding of the hinge protein to the heme subunit was essential for the non-autoxidizability of cytochrome c1 subcomplex. In addition, it was suggested that cysteinyl residues in the subcomplex must be involved to a great extent in the stable binding between the two subunits.

AB - The autoxidizability of beef heart cytochrome c1 was investigated in terms of the integrity of the binding of the hinge protein to the heme subunit. Cytochrome c1 was isolated as a subcomplex consisting of the heme subunit and the hinge protein. Treatment of the cytochrome c1 subcomplex with p-chloromercuribenzoate (pCMB) under mild conditions lessened the binding strength between the two subunits. They were dissociated on polyacrylamide gel electrophoresis (PAGE) under non-denaturing conditions, but were not separated by gel filtration chromatography. The pCMB-treated subcomplex had a slight autoxidizabiity. This was repressed to the level of the native subeomplex, when the mercurial compound bound to the subcomplex was removed by the addition of 2-mercaptoethanol. Concomitantly, the less stable binding between the subunits was apparently reversed to the native state. After pCMB treatment of the subcomplex, the heme subunit recovered from PAGE showed marked autoxidizabiity, even if it was treated with 2-mercapto-ethanol. Addition of cholate repressed the autoxidizability of the heme subunit after the removal of the mercurial compound. These results confirmed that the stable binding of the hinge protein to the heme subunit was essential for the non-autoxidizability of cytochrome c1 subcomplex. In addition, it was suggested that cysteinyl residues in the subcomplex must be involved to a great extent in the stable binding between the two subunits.

UR - http://www.scopus.com/inward/record.url?scp=0022815232&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022815232&partnerID=8YFLogxK

M3 - Article

C2 - 3029052

AN - SCOPUS:0022815232

VL - 100

SP - 1165

EP - 1173

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 5

ER -