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  • 标题:Site-specific collapse dynamics guide the formation of the cytochrome c′ four-helix bundle
  • 本地全文:下载
  • 作者:Tetsunari Kimura ; Jennifer C. Lee ; Harry B. Gray
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:2007
  • 卷号:104
  • 期号:1
  • 页码:117-122
  • DOI:10.1073/pnas.0609413103
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:The evolution of tryptophan-to-heme (W/heme) distance distributions extracted from analysis of fluorescence energy transfer kinetics during the refolding of Rhodopseudomonas palustris cytochrome c' reveals dramatic differences between two variants [W32 (Q1A/F32W/W72F) and W72 (Q1A)]. Both W32/heme and W72/heme distance distributions measured at the earliest time point attainable with a continuous-flow mixer (150 {micro}s) confirm that the polypeptide ensemble is not uniformly collapsed and that native structure is not formed. Time-resolved fluorescence spectra indicate that W32 is sequestered from the aqueous solution during the first 700 {micro}s of folding, whereas W72 remains exposed to solvent. The first moment of the W32/heme distance distribution evolves to its native value faster than that of W72, suggesting that the approach of W32 to the heme precedes that of W72.
  • 关键词:fluorescence energy transfer ; protein folding ; ultrafast mixer
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