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重组卡2 +- 研究色氨酸 - ANS FRET 的色氨酸残留物的化学修饰 - ATPase N 域
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Chemical Modification of the Tryptophan Residue in a Recombinant Ca2+-ATPase N-domain for Studying Tryptophan-ANS FRET
DOI:

12:07 min

October 09, 2021

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Chapters

  • 00:03Introduction
  • 01:15Determination (in silico) of the ANS and SERCA N‐Domain Interaction
  • 03:09Expression and Purification of the Recombinant N‐Domain
  • 03:39Monitor the Formation of the ANS‐N‐Domain Complex Based on ANS and N‐Domain Fluorescence Intensity Changes
  • 07:38N‐Domain Intrinsic Fluorescence Titration by Trp Chemical Modification with NBS
  • 08:43Titrate the NBS Modified N‐Domain with ANS by Recording Fluorescence Spectra at 25°C
  • 09:37Evidence of ANS Binding to the Chemically Modified N‐Domain by Excitation at λ = 370 nm
  • 10:32Results Overview
  • 11:30Conclusions

Summary

자동 번역

ANS 与 Ca2+- ATPase 重组 N 域绑定。荧光光谱在激发时以 295 nm 的波长显示类似 FRET 的图案。NBS 调解的 Trp 化学修饰抑制了 N 域的荧光,这导致 Trp 残留物和 ANS 之间没有能量转移 (FRET)。

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