Site-Directed Isotope Labeling and ATR-FTIR Difference Spectroscopy of Bacteriorhodopsin: The PeptiDe Carbonyl Group of Tyr 185 Is Structurally Active During the bR → N Transition

Cheryl F.C. Ludlam, Sanjay Sonar, Chan Ping Lee, Matthew A Coleman, Judith Herzfeld, Uttam L. RajBhandary, Kenneth J. Rothschild

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

The largest secondary structural change occurs in the bacteriorhodopsin (bR) photocycle during the M →-N transition. In this work site-directed isotope labeling (SDIL) and attenuated total reflection Fourier transform infrared (ATR-FTIR) difference spectroscopy were used to investigate this conformational change. L-Tyrosine containing a 13C isotope at the carbonyl carbon was selectively incorporated at Tyr 57, Tyr 147, and Tyr 185 by SDIL. This involves the cell-free expression of bR in the presence of Escherichia coli suppressor tRNATyrCUAaminoacylated with L-[l-13C]Tyr. ATR-FTIR difference spectroscopy reveals that of the 11 tyrosines, only the peptiDe carbonyl group of Tyr 185 undergoes a significant structural change during the bR → N transition. Along with other spectroscopic evidence, this result suggests that the Tyr 185-Pro 186 region of the protein is structurally active and may function as a hinge which facilitates the tilt of the cytoplasmic portion of the F-helix in bacteriorhodopsin during the M → N transition.

Original languageEnglish (US)
Pages (from-to)2-6
Number of pages5
JournalBiochemistry
Volume34
Issue number1
DOIs
StatePublished - Jan 1 1995
Externally publishedYes

Fingerprint

Isotope Labeling
Bacteriorhodopsins
Fourier Transform Infrared Spectroscopy
Isotopes
Labeling
Fourier transforms
Spectroscopy
Infrared radiation
Peptides
Tyrosine
Hinges
Workplace
Escherichia coli
Carbon
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Site-Directed Isotope Labeling and ATR-FTIR Difference Spectroscopy of Bacteriorhodopsin : The PeptiDe Carbonyl Group of Tyr 185 Is Structurally Active During the bR → N Transition. / Ludlam, Cheryl F.C.; Sonar, Sanjay; Lee, Chan Ping; Coleman, Matthew A; Herzfeld, Judith; RajBhandary, Uttam L.; Rothschild, Kenneth J.

In: Biochemistry, Vol. 34, No. 1, 01.01.1995, p. 2-6.

Research output: Contribution to journalArticle

Ludlam, Cheryl F.C. ; Sonar, Sanjay ; Lee, Chan Ping ; Coleman, Matthew A ; Herzfeld, Judith ; RajBhandary, Uttam L. ; Rothschild, Kenneth J. / Site-Directed Isotope Labeling and ATR-FTIR Difference Spectroscopy of Bacteriorhodopsin : The PeptiDe Carbonyl Group of Tyr 185 Is Structurally Active During the bR → N Transition. In: Biochemistry. 1995 ; Vol. 34, No. 1. pp. 2-6.
@article{5f335f1daff94ddba1b6f72a37d8b602,
title = "Site-Directed Isotope Labeling and ATR-FTIR Difference Spectroscopy of Bacteriorhodopsin: The PeptiDe Carbonyl Group of Tyr 185 Is Structurally Active During the bR → N Transition",
abstract = "The largest secondary structural change occurs in the bacteriorhodopsin (bR) photocycle during the M →-N transition. In this work site-directed isotope labeling (SDIL) and attenuated total reflection Fourier transform infrared (ATR-FTIR) difference spectroscopy were used to investigate this conformational change. L-Tyrosine containing a 13C isotope at the carbonyl carbon was selectively incorporated at Tyr 57, Tyr 147, and Tyr 185 by SDIL. This involves the cell-free expression of bR in the presence of Escherichia coli suppressor tRNATyrCUAaminoacylated with L-[l-13C]Tyr. ATR-FTIR difference spectroscopy reveals that of the 11 tyrosines, only the peptiDe carbonyl group of Tyr 185 undergoes a significant structural change during the bR → N transition. Along with other spectroscopic evidence, this result suggests that the Tyr 185-Pro 186 region of the protein is structurally active and may function as a hinge which facilitates the tilt of the cytoplasmic portion of the F-helix in bacteriorhodopsin during the M → N transition.",
author = "Ludlam, {Cheryl F.C.} and Sanjay Sonar and Lee, {Chan Ping} and Coleman, {Matthew A} and Judith Herzfeld and RajBhandary, {Uttam L.} and Rothschild, {Kenneth J.}",
year = "1995",
month = "1",
day = "1",
doi = "10.1021/bi00001a001",
language = "English (US)",
volume = "34",
pages = "2--6",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "1",

}

TY - JOUR

T1 - Site-Directed Isotope Labeling and ATR-FTIR Difference Spectroscopy of Bacteriorhodopsin

T2 - The PeptiDe Carbonyl Group of Tyr 185 Is Structurally Active During the bR → N Transition

AU - Ludlam, Cheryl F.C.

AU - Sonar, Sanjay

AU - Lee, Chan Ping

AU - Coleman, Matthew A

AU - Herzfeld, Judith

AU - RajBhandary, Uttam L.

AU - Rothschild, Kenneth J.

PY - 1995/1/1

Y1 - 1995/1/1

N2 - The largest secondary structural change occurs in the bacteriorhodopsin (bR) photocycle during the M →-N transition. In this work site-directed isotope labeling (SDIL) and attenuated total reflection Fourier transform infrared (ATR-FTIR) difference spectroscopy were used to investigate this conformational change. L-Tyrosine containing a 13C isotope at the carbonyl carbon was selectively incorporated at Tyr 57, Tyr 147, and Tyr 185 by SDIL. This involves the cell-free expression of bR in the presence of Escherichia coli suppressor tRNATyrCUAaminoacylated with L-[l-13C]Tyr. ATR-FTIR difference spectroscopy reveals that of the 11 tyrosines, only the peptiDe carbonyl group of Tyr 185 undergoes a significant structural change during the bR → N transition. Along with other spectroscopic evidence, this result suggests that the Tyr 185-Pro 186 region of the protein is structurally active and may function as a hinge which facilitates the tilt of the cytoplasmic portion of the F-helix in bacteriorhodopsin during the M → N transition.

AB - The largest secondary structural change occurs in the bacteriorhodopsin (bR) photocycle during the M →-N transition. In this work site-directed isotope labeling (SDIL) and attenuated total reflection Fourier transform infrared (ATR-FTIR) difference spectroscopy were used to investigate this conformational change. L-Tyrosine containing a 13C isotope at the carbonyl carbon was selectively incorporated at Tyr 57, Tyr 147, and Tyr 185 by SDIL. This involves the cell-free expression of bR in the presence of Escherichia coli suppressor tRNATyrCUAaminoacylated with L-[l-13C]Tyr. ATR-FTIR difference spectroscopy reveals that of the 11 tyrosines, only the peptiDe carbonyl group of Tyr 185 undergoes a significant structural change during the bR → N transition. Along with other spectroscopic evidence, this result suggests that the Tyr 185-Pro 186 region of the protein is structurally active and may function as a hinge which facilitates the tilt of the cytoplasmic portion of the F-helix in bacteriorhodopsin during the M → N transition.

UR - http://www.scopus.com/inward/record.url?scp=33751155072&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33751155072&partnerID=8YFLogxK

U2 - 10.1021/bi00001a001

DO - 10.1021/bi00001a001

M3 - Article

C2 - 7819197

AN - SCOPUS:0028988375

VL - 34

SP - 2

EP - 6

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 1

ER -