Protein conformational changes during the bacteriorhodopsin photocycle: A fourier transform infrared/resonance Raman study of the alkaline form of the mutant Asp-85 → Asn

Anders Nilsson, Parshuram Rath, Jerzy Olejnik, Matthew A Coleman, Kenneth J. Rothschild

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Bacteriorhodopsin is a light-driven proton pump, which undergoes a photocycle consisting of several distinct intermediates. Previous studies have established that the M → N step of this photocycle involves a major conformational change of membrane embedded α-helices. In order to further investigate this conformational change, we have studied the photocycle of the high pH form of the mutant Asp-85 → Asn (D85N(alk)). In contrast to wild type bacteriorhodopsin, D85N(alk) has a deprotonated Schiff base and a blue- shifted absorption near 410 nm, yet it still transports protons in the same direction as wild type bacteriorhodopsin (Tittor, J., Schweiger, U., Ocsterhelt, D. and Barnberg, E. (1994) Biophys. J., 67, 1682-1690). Resonance Raman spectroscopy of D85N(alk) and D85N(alk) regenerated with retinal labeled at the C-15 position with deuterium reveals the existence of an all- trans configuration of the chromophore. Fourier transform infrared difference spectroscopy shows that the photocycle of this light-adapted form involves similar events as the wild type bacteriorhodopsin photocycle including the M → N protein conformational change. These results help to explain the ability of D85N(alk) to transport protons and demonstrate that the M → N conformational change can occur even in the photocycle of an unprotonated Schiff base form of bacteriorhodopsin.

Original languageEnglish (US)
Pages (from-to)29746-29751
Number of pages6
JournalJournal of Biological Chemistry
Volume270
Issue number50
DOIs
StatePublished - Dec 15 1995
Externally publishedYes

Fingerprint

Bacteriorhodopsins
Fourier Analysis
Fourier transforms
Infrared radiation
Schiff Bases
Proteins
Protons
Light
Proton Pumps
Raman Spectrum Analysis
Deuterium
Fourier Transform Infrared Spectroscopy
Chromophores
Raman spectroscopy
Spectroscopy
Membranes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Protein conformational changes during the bacteriorhodopsin photocycle : A fourier transform infrared/resonance Raman study of the alkaline form of the mutant Asp-85 → Asn. / Nilsson, Anders; Rath, Parshuram; Olejnik, Jerzy; Coleman, Matthew A; Rothschild, Kenneth J.

In: Journal of Biological Chemistry, Vol. 270, No. 50, 15.12.1995, p. 29746-29751.

Research output: Contribution to journalArticle

@article{901056c7bd5c40fea0683eda2acaf0b2,
title = "Protein conformational changes during the bacteriorhodopsin photocycle: A fourier transform infrared/resonance Raman study of the alkaline form of the mutant Asp-85 → Asn",
abstract = "Bacteriorhodopsin is a light-driven proton pump, which undergoes a photocycle consisting of several distinct intermediates. Previous studies have established that the M → N step of this photocycle involves a major conformational change of membrane embedded α-helices. In order to further investigate this conformational change, we have studied the photocycle of the high pH form of the mutant Asp-85 → Asn (D85N(alk)). In contrast to wild type bacteriorhodopsin, D85N(alk) has a deprotonated Schiff base and a blue- shifted absorption near 410 nm, yet it still transports protons in the same direction as wild type bacteriorhodopsin (Tittor, J., Schweiger, U., Ocsterhelt, D. and Barnberg, E. (1994) Biophys. J., 67, 1682-1690). Resonance Raman spectroscopy of D85N(alk) and D85N(alk) regenerated with retinal labeled at the C-15 position with deuterium reveals the existence of an all- trans configuration of the chromophore. Fourier transform infrared difference spectroscopy shows that the photocycle of this light-adapted form involves similar events as the wild type bacteriorhodopsin photocycle including the M → N protein conformational change. These results help to explain the ability of D85N(alk) to transport protons and demonstrate that the M → N conformational change can occur even in the photocycle of an unprotonated Schiff base form of bacteriorhodopsin.",
author = "Anders Nilsson and Parshuram Rath and Jerzy Olejnik and Coleman, {Matthew A} and Rothschild, {Kenneth J.}",
year = "1995",
month = "12",
day = "15",
doi = "10.1074/jbc.270.50.29746",
language = "English (US)",
volume = "270",
pages = "29746--29751",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "50",

}

TY - JOUR

T1 - Protein conformational changes during the bacteriorhodopsin photocycle

T2 - A fourier transform infrared/resonance Raman study of the alkaline form of the mutant Asp-85 → Asn

AU - Nilsson, Anders

AU - Rath, Parshuram

AU - Olejnik, Jerzy

AU - Coleman, Matthew A

AU - Rothschild, Kenneth J.

PY - 1995/12/15

Y1 - 1995/12/15

N2 - Bacteriorhodopsin is a light-driven proton pump, which undergoes a photocycle consisting of several distinct intermediates. Previous studies have established that the M → N step of this photocycle involves a major conformational change of membrane embedded α-helices. In order to further investigate this conformational change, we have studied the photocycle of the high pH form of the mutant Asp-85 → Asn (D85N(alk)). In contrast to wild type bacteriorhodopsin, D85N(alk) has a deprotonated Schiff base and a blue- shifted absorption near 410 nm, yet it still transports protons in the same direction as wild type bacteriorhodopsin (Tittor, J., Schweiger, U., Ocsterhelt, D. and Barnberg, E. (1994) Biophys. J., 67, 1682-1690). Resonance Raman spectroscopy of D85N(alk) and D85N(alk) regenerated with retinal labeled at the C-15 position with deuterium reveals the existence of an all- trans configuration of the chromophore. Fourier transform infrared difference spectroscopy shows that the photocycle of this light-adapted form involves similar events as the wild type bacteriorhodopsin photocycle including the M → N protein conformational change. These results help to explain the ability of D85N(alk) to transport protons and demonstrate that the M → N conformational change can occur even in the photocycle of an unprotonated Schiff base form of bacteriorhodopsin.

AB - Bacteriorhodopsin is a light-driven proton pump, which undergoes a photocycle consisting of several distinct intermediates. Previous studies have established that the M → N step of this photocycle involves a major conformational change of membrane embedded α-helices. In order to further investigate this conformational change, we have studied the photocycle of the high pH form of the mutant Asp-85 → Asn (D85N(alk)). In contrast to wild type bacteriorhodopsin, D85N(alk) has a deprotonated Schiff base and a blue- shifted absorption near 410 nm, yet it still transports protons in the same direction as wild type bacteriorhodopsin (Tittor, J., Schweiger, U., Ocsterhelt, D. and Barnberg, E. (1994) Biophys. J., 67, 1682-1690). Resonance Raman spectroscopy of D85N(alk) and D85N(alk) regenerated with retinal labeled at the C-15 position with deuterium reveals the existence of an all- trans configuration of the chromophore. Fourier transform infrared difference spectroscopy shows that the photocycle of this light-adapted form involves similar events as the wild type bacteriorhodopsin photocycle including the M → N protein conformational change. These results help to explain the ability of D85N(alk) to transport protons and demonstrate that the M → N conformational change can occur even in the photocycle of an unprotonated Schiff base form of bacteriorhodopsin.

UR - http://www.scopus.com/inward/record.url?scp=0029620915&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029620915&partnerID=8YFLogxK

U2 - 10.1074/jbc.270.50.29746

DO - 10.1074/jbc.270.50.29746

M3 - Article

C2 - 8530365

AN - SCOPUS:0029620915

VL - 270

SP - 29746

EP - 29751

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 50

ER -