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

Biochemistry
F Aladar BencsathS R Goodman

Abstract

We have previously demonstrated that the membrane skeletons of irreversibly sickled cells (ISCs) dissociate more slowly at 37 degrees 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 beta-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 beta-actin. We synthesized the associated ISC beta-actin tryptic cystine-peptide (KCF-CDVDIR), characterized it by HPLC, MS. and MSMS, and identified it in the tryptic digest of the ISC beta-actin. These results support our earlier suggestion that the oxidative change in ISC beta-actin is a major cause of the irreversible sickling phenomenon.

Citations

Jan 8, 2005·Chembiochem : a European Journal of Chemical Biology·Michael Famulok, Günter Mayer
Jun 2, 1998·Free Radical Biology & Medicine·P BrowneR P Hebbel
Dec 19, 2009·Cell Death and Differentiation·J E LeadshamC W Gourlay
Aug 12, 2004·Molecular Biology of the Cell·Brian K Haarer, David C Amberg
Feb 9, 2007·Molecular Biology of the Cell·Michelle E Farah, David C Amberg
Oct 31, 2012·Molecular Aspects of Medicine·Niki ChondrogianniEfstathios S Gonos
Mar 11, 2009·Free Radical Biology & Medicine·Wei Du, Kathryn R Ayscough
Jun 3, 2011·Cytoskeleton·Michelle E FarahDavid C Amberg
May 21, 2015·Brain Structure & Function·Ria UhlemannGolo Kronenberg
Feb 26, 2008·Biochimica Et Biophysica Acta·Jane E Leadsham, Campbell W Gourlay
Jun 1, 1997·American Journal of Hematology·A ShartavaS R Goodman
Jul 17, 2013·Experimental Biology and Medicine·Steven R GoodmanMarko Zivanic
Aug 19, 2015·Experimental Biology and Medicine·Steven R GoodmanWarren E Zimmer
Aug 21, 2013·Applied and Environmental Microbiology·Nuria Pujol-CarrionMaria Angeles de la Torre-Ruiz
Sep 5, 2019·Experimental Biology and Medicine·Steven R GoodmanDavid Kakhniashvili
Jul 11, 2008·The Biochemical Journal·Vernonica E Franklin-Tong, Campbell W Gourlay

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