title: A New Method to Determine the Transmembrane Conformation of Substrates in Intramembrane Proteolysis by Deep-UV Resonance Raman Spectroscopy.
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.
authors.affiliation: University of Missouri, Columbia, MO, United States.
authors.forename: J W
authors.initials: JW
authors.lastname: Cooley
authors.affiliation: Icahn School of Medicine at Mount Sinai, New York, NY, United States.
authors.forename: A
authors.initials: A
authors.lastname: Abdine
authors.affiliation: University of Missouri, Columbia, MO, United States.
authors.forename: M
authors.initials: M
authors.lastname: Brown
authors.affiliation: Icahn School of Medicine at Mount Sinai, New York, NY, United States.
authors.forename: J
authors.initials: J
authors.lastname: Chavez
authors.affiliation: University of Missouri, Columbia, MO, United States.
authors.forename: B
authors.initials: B
authors.lastname: Lada
authors.affiliation: University of Missouri, Columbia, MO, United States.
authors.forename: R D
authors.initials: RD
authors.lastname: JiJi
authors.affiliation: Icahn School of Medicine at Mount Sinai, New York, NY, United States. Electronic address: iban.ubarretxena@mssm.edu.
authors.forename: I
authors.initials: I
authors.lastname: Ubarretxena-Belandia
mesh_terms: D002462:Cell Membrane / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry; D008967:Molecular Biology / Q000379:methods*; D011487:Protein Conformation; D059748:Proteolysis; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013379:Substrate Specificity
chemical_list: D008565:Membrane Proteins
keywords: Intramembrane proteolysis; Raman spectroscopy; Rhomboid; Transmembrane domain conformation