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A New Method to Determine the Transmembrane Conformation of Substrates in Intramembrane Proteolysis by Deep-UV Resonance Raman Spectroscopy

  • J. W. Cooley
  • , A. Abdine
  • , M. Brown
  • , J. Chavez
  • , B. Lada
  • , R. D. JiJi
  • , I. Ubarretxena-Belandia

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

2 Scopus citations

Abstract

We present a new method based on deep-UV resonance Raman spectroscopy to determine the backbone conformation of intramembrane protease substrates. The classical amide vibrational modes reporting on the conformation of just the transmembrane region of the substrate can be resolved from solvent exchangeable regions outside the detergent micelle by partial deuteration of the solvent. In the presence of isotopically triple-labeled intramembrane protease, these amide modes can be accurately measured to monitor the transmembrane conformation of the substrate during intramembrane proteolysis.

Original languageEnglish
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages207-228
Number of pages22
DOIs
StatePublished - 2017

Publication series

NameMethods in Enzymology
Volume584
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Intramembrane proteolysis
  • Raman spectroscopy
  • Rhomboid
  • Transmembrane domain conformation

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