Crosslinking of troponin complex with 1,3-difluoro-4,6-dinitrobenzene. Identification of the crosslink formed between troponin C and troponin I in the absence of Ca2+

Anatoly B. Dobrovol'sky, Nikolay B. Gusev, P. Friedrich

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The single SH-group of rabbit skeletal muscle troponin C (Cys-98) was reacted with the bifunctional reagent, 1,3-difluoro-4,6-dinitrobenzene. This labelled troponin C was used to reconstitute the troponin complex by the addition of equimolar amounts of troponin T and troponin I. The second function of the bifunctional reagent was triggered in the complex by an increase of pH. A crosslink was formed between troponin C and troponin I both in the presence and absence of Ca2+, but the probability of crosslinking was decreased by Ca2+. Covalently linked troponin C-trophonin I was isolated from the complex crosslinked without Ca2+, and cleaved by CNBr. The analysis of crosslinked peptides has revealed that in the presence of Mg2+ and absence of Ca2+ the crosslink in the troponin complex is formed between Cys-98 of troponin C and Cys-133 of troponin I.

Original languageEnglish
Pages (from-to)144-151
Number of pages8
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume789
Issue number2
DOIs
Publication statusPublished - Sep 11 1984

Fingerprint

Troponin C
Troponin
Troponin I
Crosslinking
Cross-Linking Reagents
Troponin T
Muscle
Skeletal Muscle
1,5-difluoro-2,4-dinitrobenzene
Rabbits
Peptides

Keywords

  • (Rabbit skeletal muscle)
  • Peptide analysis
  • Protein crosslinking
  • Reconstitution
  • Troponin

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Structural Biology

Cite this

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title = "Crosslinking of troponin complex with 1,3-difluoro-4,6-dinitrobenzene. Identification of the crosslink formed between troponin C and troponin I in the absence of Ca2+",
abstract = "The single SH-group of rabbit skeletal muscle troponin C (Cys-98) was reacted with the bifunctional reagent, 1,3-difluoro-4,6-dinitrobenzene. This labelled troponin C was used to reconstitute the troponin complex by the addition of equimolar amounts of troponin T and troponin I. The second function of the bifunctional reagent was triggered in the complex by an increase of pH. A crosslink was formed between troponin C and troponin I both in the presence and absence of Ca2+, but the probability of crosslinking was decreased by Ca2+. Covalently linked troponin C-trophonin I was isolated from the complex crosslinked without Ca2+, and cleaved by CNBr. The analysis of crosslinked peptides has revealed that in the presence of Mg2+ and absence of Ca2+ the crosslink in the troponin complex is formed between Cys-98 of troponin C and Cys-133 of troponin I.",
keywords = "(Rabbit skeletal muscle), Peptide analysis, Protein crosslinking, Reconstitution, Troponin",
author = "Dobrovol'sky, {Anatoly B.} and Gusev, {Nikolay B.} and P. Friedrich",
year = "1984",
month = "9",
day = "11",
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language = "English",
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T1 - Crosslinking of troponin complex with 1,3-difluoro-4,6-dinitrobenzene. Identification of the crosslink formed between troponin C and troponin I in the absence of Ca2+

AU - Dobrovol'sky, Anatoly B.

AU - Gusev, Nikolay B.

AU - Friedrich, P.

PY - 1984/9/11

Y1 - 1984/9/11

N2 - The single SH-group of rabbit skeletal muscle troponin C (Cys-98) was reacted with the bifunctional reagent, 1,3-difluoro-4,6-dinitrobenzene. This labelled troponin C was used to reconstitute the troponin complex by the addition of equimolar amounts of troponin T and troponin I. The second function of the bifunctional reagent was triggered in the complex by an increase of pH. A crosslink was formed between troponin C and troponin I both in the presence and absence of Ca2+, but the probability of crosslinking was decreased by Ca2+. Covalently linked troponin C-trophonin I was isolated from the complex crosslinked without Ca2+, and cleaved by CNBr. The analysis of crosslinked peptides has revealed that in the presence of Mg2+ and absence of Ca2+ the crosslink in the troponin complex is formed between Cys-98 of troponin C and Cys-133 of troponin I.

AB - The single SH-group of rabbit skeletal muscle troponin C (Cys-98) was reacted with the bifunctional reagent, 1,3-difluoro-4,6-dinitrobenzene. This labelled troponin C was used to reconstitute the troponin complex by the addition of equimolar amounts of troponin T and troponin I. The second function of the bifunctional reagent was triggered in the complex by an increase of pH. A crosslink was formed between troponin C and troponin I both in the presence and absence of Ca2+, but the probability of crosslinking was decreased by Ca2+. Covalently linked troponin C-trophonin I was isolated from the complex crosslinked without Ca2+, and cleaved by CNBr. The analysis of crosslinked peptides has revealed that in the presence of Mg2+ and absence of Ca2+ the crosslink in the troponin complex is formed between Cys-98 of troponin C and Cys-133 of troponin I.

KW - (Rabbit skeletal muscle)

KW - Peptide analysis

KW - Protein crosslinking

KW - Reconstitution

KW - Troponin

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