Journal
/
/
트립토판-ANS FRET 연구를 위한 재조합 Ca2+-ATPase N-도메인에서 트립토판 잔류물의 화학적 수정
JoVE Journal
Biochemistry
Author Produced
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
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

,

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

Automatic Translation

ANS는 Ca2+-ATPase 재조합 N-도메인에 바인딩합니다. 형광 스펙트럼은 295 nm의 파장에서 여기시 면에 FRET 와 같은 패턴을 표시합니다. Trp의 NBS 중재 화학 적 변형은 Trp 잔류물과 ANS 사이의 에너지 전송 (FRET)의 부재로 이어지는 N 도메인의 형광을 담금질합니다.

Related Videos

Read Article