期刊
FREE RADICAL BIOLOGY AND MEDICINE
卷 51, 期 3, 页码 755-762出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2011.05.021
关键词
alpha B-crystallin; Copper binding; Redox attenuation; Small-angle X-ray scattering; Free radicals
资金
- Council of Scientific and Industrial Research, New Delhi
- Department of Science and Technology, India
Oxidative stress, Cu2+ homeostasis, and small heat shock proteins (sHsp's) have important implications in several neurodegenerative diseases. The ubiquitous sHsp alpha B-crystallin is an oligomeric protein that binds Cu2+ We have investigated the relative contributions of the N- and C-terminal (C-TD alpha B-crystallin) domains of alpha B-crystallin to its Cu2+-binding and redox-attenuation properties and mapped the Cu2+-binding regions. C-TD alpha B-crystallin binds Cu2+ with slightly less affinity and inhibits Cu2+-catalyzed, ascorbate-mediated generation of ROS to a lesser extent than alpha B-crystallin. [Cu2+]/[subunit] stoichiometries for redox attenuation by alpha B-crystallin and C-TD alpha B-crystallin are 5 and 2, respectively. Both alpha B-crystallin and C-TD alpha B-crystallin also inhibit the Fenton reaction of hydroxyl radical formation. Trypsinization of alpha B-crystallin bound to a Cu2+-NTA column and MALDI-TOF analysis of column-bound peptides yielded three peptides located in the N-terminal domain, and in-solution trypsinization of alpha B-crystallin followed by Cu2+-NTA column chromatography identified four additional Cu2+-binding peptides located in the C-terminal domain. Thus, Cu2+-binding regions are distributed in the N- and C-terminal domains. Small-angle X-ray scattering and sedimentation-velocity measurements indicate quaternary structural changes in alpha B-crystallin upon Cu2+ binding. Our study indicates that an oligomer of alpha B-crystallin can sequester a large number (similar to 150) of Cu2+ ions. It acts like a Cu2+ sponge, exhibits redox attenuation of Cu2+, and has potential roles in Cu2+ homeostasis and in preventing oxidative stress. (C) 2011 Elsevier Inc. All rights reserved.
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