Inhibition of Hsp72-mediated protein refolding by 4-hydroxy-2-nonenal

被引:94
作者
Carbone, DL [1 ]
Doorn, JA [1 ]
Kiebler, Z [1 ]
Sampey, BP [1 ]
Petersen, DR [1 ]
机构
[1] Univ Colorado, Hlth Sci Ctr, Sch Pharm, Dept Pharmaceut Sci, Denver, CO 80262 USA
关键词
D O I
10.1021/tx049838g
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
A proteomic approach was applied to liver cytosol from rats fed a diet consisting of high fat and ethanol to identify 4-hydroxy-2-nonenal (4-HNE)-modified proteins in vivo. Cytosolic Hsp72, the inducible variant of the Hsp70 heat shock protein family, was consistently among the proteins modified by 4-HNE. Despite 1.3-fold induction of Hsp72 in the livers of ethanol-fed animals, no increase in Hsp70-mediated luciferase refolding in isolated heptocytes was observed, suggesting inhibition of this process by 4-HNE. A 50% and 75% reduction in luciferase refolding efficiency was observed in rabbit reticulocyte lysate (RRL) supplemented with recombinant Hsp72 which had been modified in vitro with 10 and 100 muM 4-HNE, respectively. This observation was accompanied by a 25% and 50% decrease in substrate binding by the chaperone following the same treatment; however, no effect on complex formation between Hsp72 and its co-chaperone Hsp40 was observed. Trypsin digest and mass spectral analysis of Hsp72 treated with 10 and 100 muM 4-HNE consistently identified adduct formation at Cys267 in the ATPase domain of the chaperone. The role of this residue in the observed inhibition was demonstrated through the use of DnaK, a bacterial Hsp70 variant lacking Cys267. DnaK was resistant to 4-HNE inactivation. Additionally, Hsp72 was resistant to inactivation by the thiol-unreactive aldehyde malondialdehyde (MDA), further supporting a role for Cys in Hsp72 inhibition by 4-HNE. Finally, the affinity of Hsp72 for ATP was decreased 32% and 72% following treatment of the chaperone with 10 and 100 muM 4-HNE, respectively. In a model of chronic alcoholic liver injury, induction of Hsp72 was not accompanied by an increase in protein refolding ability. This is likely the result of 4-HNE modification of the Hsp72 ATPase domain.
引用
收藏
页码:1459 / 1467
页数:9
相关论文
共 39 条
[1]   Induction of redox instability of bovine myoglobin by adduction with 4-hydroxy-2-nonenal [J].
Alderton, AL ;
Faustman, C ;
Liebler, DC ;
Hill, DW .
BIOCHEMISTRY, 2003, 42 (15) :4398-4405
[2]   Interaction of the molecular chaperone Hsp70 with human NAD(P)H:quinone oxidoreductase 1 [J].
Anwar, A ;
Siegel, D ;
Kepa, JK ;
Ross, D .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (16) :14060-14067
[3]   Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome [J].
Beere, HM ;
Wolf, BB ;
Cain, K ;
Mosser, DD ;
Mahboubi, A ;
Kuwana, T ;
Tailor, P ;
Morimoto, RI ;
Cohen, GM ;
Green, DR .
NATURE CELL BIOLOGY, 2000, 2 (08) :469-475
[4]  
Bercovich B, 1997, J BIOL CHEM, V272, P9002
[5]   Stress proteins and sh-groups in oxidant-induced cell damage after acute ethanol administration in rat [J].
Calabrese, V ;
Renis, M ;
Calderone, A ;
Russo, A ;
Barcellona, ML ;
Rizza, V .
FREE RADICAL BIOLOGY AND MEDICINE, 1996, 20 (03) :391-397
[6]  
Diehl AM, 2001, ALCOHOL CLIN EXP RES, V25, p8S, DOI 10.1097/00000374-200105051-00004
[7]   Peptidase activities of the multicatalytic protease in rat liver after voluntary and intragastric ethanol administration [J].
Donohue, TM ;
Zetterman, RK ;
Zhang-Gouillon, ZQ ;
French, SW .
HEPATOLOGY, 1998, 28 (02) :486-491
[8]   Covalent modification of amino acid nucleophiles by the lipid peroxidation products 4-hydroxy-2-nonenal and 4-oxo-2-nonenal [J].
Doorn, JA ;
Petersen, DR .
CHEMICAL RESEARCH IN TOXICOLOGY, 2002, 15 (11) :1445-1450
[9]   CHEMISTRY AND BIOCHEMISTRY OF 4-HYDROXYNONENAL, MALONALDEHYDE AND RELATED ALDEHYDES [J].
ESTERBAUER, H ;
SCHAUR, RJ ;
ZOLLNER, H .
FREE RADICAL BIOLOGY AND MEDICINE, 1991, 11 (01) :81-128
[10]  
ESTERBAUER H, 1989, MEMBRANE LIPID OXIDA, V1, P239