https://scholars.lib.ntu.edu.tw/handle/123456789/406740
DC 欄位 | 值 | 語言 |
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dc.contributor.author | Wang S.S.-S. | en_US |
dc.contributor.author | Wen W.-S. | en_US |
dc.creator | Wen W.-S.;Wang S.S.-S. | - |
dc.date.accessioned | 2019-05-09T04:08:32Z | - |
dc.date.available | 2019-05-09T04:08:32Z | - |
dc.date.issued | 2010 | - |
dc.identifier.issn | 10900535 | - |
dc.identifier.uri | https://scholars.lib.ntu.edu.tw/handle/123456789/406740 | - |
dc.description.abstract | Purpose: Human £^D crystallin is a principal protein component of the human eye lens and associated with the development of juvenile and mature-onset cataracts. Exposure to ultraviolet (UV) light is thought to perturb protein structure and eventually lead to aggregation. This work is aimed at exploring the effects of UV-C irradiation on recombinant human £^D-crystallin (HGDC). Methods: Recombinant HGDC proteins were expressed in E. coli strain BL21(DE3) harboring plasmid pEHisHGDC and purified using chromatographic methods. The proteins were then exposed to UV-C light (£fmax=254 nm, 15 W) at the intensity of 420, 800, or 1850 £gW/cm2. The UV-C-unexposed, supernatant fraction of UV-C-exposed, and re-dissolved precipitated fraction of UV-C exposed preparations were characterized by SDS-PAGE, turbidity measurement, CD spectroscopy, tryptophan fluorescence spectroscopy, acrylamide fluorescence quenching analysis, and sulfhydryl group measurements. Results: The turbidity of the HGDC sample solution was found to be positively correlated with HGDC concentration, UV-C irradiation intensity, and UV-C irradiation duration. When exposed to UV-C, HGDC sample solutions became visibly turbid and a noticeable amount of larger protein particle, perceptible to the naked eye, was observed upon prolonged irradiation. The precipitated fraction of irradiated HGDC sample was found to be re-dissolved by guanidine hydrochloride. Electrophoresis, acrylamide fluorescence quenching, and spectroscopic analyses revealed differences in structures among the non-irradiated HGDC, the supernatant fraction of irradiated HGDC, and the re-dissolved precipitated fraction of irradiated HGDC. Through the use of L-cysteine, the measurements of sulfhydryl contents, and the reducing as well as non-reducing SDS-PAGE, our data further suggested that disulfide bond formation and/or cleavage probably play an important role in aggregation and/or precipitation of HGDC elicited by UV-C irradiation. Conclusions: Our findings highlight the close connections among disulfide bond cleavage and/or formation, intermolecular interactions, and the resultant formation of aggregates of HGDC induced by UV-C irradiation. The results from this research may not only contribute to the understanding of the environmental factors causing protein aggregation but also have implications for deciphering the molecular mechanism of cataractogenesis. ? 2010 Molecular Vision. | - |
dc.language | en | - |
dc.relation.ispartof | Molecular Vision | en_US |
dc.title | Examining the influence of ultraviolet C irradiation on recombinant human £^D-crystallin | en_US |
dc.type | journal article | en |
dc.identifier.scopus | 2-s2.0-78650762942 | - |
dc.identifier.url | https://www.scopus.com/inward/record.uri?eid=2-s2.0-78650762942&partnerID=40&md5=aa14a3725f0dc7a696719e947c3f2bda | - |
dc.relation.pages | 2777-2790 | - |
dc.relation.journalvolume | 16 | - |
item.cerifentitytype | Publications | - |
item.openairetype | journal article | - |
item.fulltext | no fulltext | - |
item.grantfulltext | none | - |
item.openairecristype | http://purl.org/coar/resource_type/c_6501 | - |
crisitem.author.dept | Chemical Engineering | - |
crisitem.author.orcid | exist | - |
crisitem.author.parentorg | College of Engineering | - |
顯示於: | 化學工程學系 |
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