OXIDATIVE STRESS TRANSFORMS ACETYLCHOLINESTERASE TO A MOLTEN-GLOBULE-LIKE STATE

被引:55
作者
WEINER, L [1 ]
KREIMER, D [1 ]
ROTH, E [1 ]
SILMAN, I [1 ]
机构
[1] WEIZMANN INST SCI,DEPT NEUROBIOL,IL-76100 REHOVOT,ISRAEL
关键词
D O I
10.1006/bbrc.1994.1130
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Exposure of purified acetylcholinesterase from Torpedo californica to a system generating oxygen radicals (viz. ascorbic acid/Fe(EDTA)2/H2O2) inactivated the enzyme. The enzyme retained its native dimeric form, but electrophoresis under denaturing conditions showed some cleavage of peptide bonds. Spectroscopic characterization revealed a shift to the red in the intrinsic fluorescence emission peak, a large decrease in molar ellipticity in the near UV with a much smaller decrease in the far UV, and increased binding of the amphiphilic probe, 1-anilino-8-naphthalene sulfonate, all relative to native enzyme. The treated enzyme was also highly susceptible to proteolysis. These data show that oxygen radical treatment converts acetylcholinesterase to a partially unfolded state, which retains most of its secondary structure but lacks substantial tertiary structure, thus resembling a ‘molten globule’ state. This model system may offer a mechanism for explaining the consequences of oxidative stress in vivo: partially unfolded proteins generated by oxidative stress may interact with molecular chaperons of the heat shock family, thus activating the heat-shock factor and, thereby, activating heat-shock genes. © 1994 Academic Press, Inc.
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收藏
页码:915 / 922
页数:8
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