Identification of the disulfide-linked peptide in irreversibly sickled cell β-actin

F. Aladar Bencsath, Archil Shartava, Carlos A. Monteiro, Steven Goodman

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

We have previously demonstrated that the membrane skeletons of irreversibly sickled cells (ISCs) dissociate more slowly at 37 °C, in high ionic strength Triton X-100 buffer, than do the membrane skeletons of reversibly sickled cells or control erythrocytes [Shartava et al. (1995) J. Cell. Biol. 128, 805-818]. Furthermore, we demonstrated that the major cause of this slow dissociation was a single posttranslational modification in ISC β-actin. Two sulfhydryl groups (Cys284 and Cys373) became inaccessible to thiol reagents because of this modification. We suggested the possibility that the modification was a disulfide bridge between Cys284 and Cys373 since the reducing agent dithiothreitol restored the sulfhydryl groups. In this article, we directly demonstrate the existence of the disulfide bridge between cysteine284 and cysteine373 in ISC β-actin. We synthesized the associated ISC β-actin tryptic cystine-peptide (KCF-CDVDIR), characterized it by HPLC, MS, and MSMS, and identified it in the tryptic digest of the ISC β-actin. These results support our earlier suggestion that the oxidative change in ISC β-actin is a major cause of the irreversible sickling phenomenon.

Original languageEnglish (US)
Pages (from-to)4403-4408
Number of pages6
JournalBiochemistry
Volume35
Issue number14
DOIs
StatePublished - Apr 9 1996
Externally publishedYes

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Disulfides
Actins
Peptides
Membranes
Sulfhydryl Reagents
Skeleton
Cystine
Dithiothreitol
Reducing Agents
Octoxynol
Ionic strength
Buffers
Post Translational Protein Processing
Osmolar Concentration
Erythrocytes
High Pressure Liquid Chromatography

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Identification of the disulfide-linked peptide in irreversibly sickled cell β-actin. / Bencsath, F. Aladar; Shartava, Archil; Monteiro, Carlos A.; Goodman, Steven.

In: Biochemistry, Vol. 35, No. 14, 09.04.1996, p. 4403-4408.

Research output: Contribution to journalArticle

Bencsath, F. Aladar ; Shartava, Archil ; Monteiro, Carlos A. ; Goodman, Steven. / Identification of the disulfide-linked peptide in irreversibly sickled cell β-actin. In: Biochemistry. 1996 ; Vol. 35, No. 14. pp. 4403-4408.
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