Formation of a new buried charge drives a large-amplitude protein quake in photoreceptor activation

A. Xie, L. Kelemen, J. Hendriks, B. J. White, K. J. Hellingwerf, W. D. Hoff

Research output: Contribution to journalArticle

197 Citations (Scopus)

Abstract

Photoactive yellow protein (PYP) is a eubacterial photoreceptor and a structural prototype of the PAS domain superfamily of receptor and regulatory proteins. We investigate the activation mechanism of PYP using time-resolved Fourier transform infrared (FTIR) spectroscopy. Our data provide structural, kinetic, and energetic evidence that the putative signaling state of PYP is formed during a large-amplitude protein quake that is driven by the formation of a new buried charge, COO- of the conserved Glu46, in a highly hydrophobic pocket at the active site. A protein quake is a process consisting of global conformational changes that are triggered and driven by a local structural "fault". We show that large, global structural changes take place after Glu46 ionization via intramolecular proton transfer to the anionic p-coumarate chromophore, and are suppressed by the absence of COO- formation in the E46Q mutant. Our results demonstrate the significance of buried charge formation in photoreceptor activation. This mechanism may serve as one of the general themes in activation of a range of receptor proteins. In addition, we report the results of time-resolved FTIR spectroscopy of PYP crystals. The direct comparison of time-resolved FTIR spectroscopic data of PYP in aqueous solution and in crystals reveals that the structure of the putative signaling state is not developed in P63 crystals. Therefore, when the structural developments during the functional process of a protein are experimentally determined to be very different in crystals and solutions, one must be cautious in drawing conclusions regarding the functional mechanism of proteins based on time-resolved X-ray crystallography.

Original languageEnglish
Pages (from-to)1510-1517
Number of pages8
JournalBiochemistry
Volume40
Issue number6
DOIs
Publication statusPublished - Feb 13 2001

Fingerprint

Chemical activation
Proteins
Crystals
Fourier Transform Infrared Spectroscopy
Proton transfer
X ray crystallography
X Ray Crystallography
Fourier Analysis
Chromophores
Ionization
Protons
Catalytic Domain
Fourier transforms
Infrared radiation
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Formation of a new buried charge drives a large-amplitude protein quake in photoreceptor activation. / Xie, A.; Kelemen, L.; Hendriks, J.; White, B. J.; Hellingwerf, K. J.; Hoff, W. D.

In: Biochemistry, Vol. 40, No. 6, 13.02.2001, p. 1510-1517.

Research output: Contribution to journalArticle

Xie, A. ; Kelemen, L. ; Hendriks, J. ; White, B. J. ; Hellingwerf, K. J. ; Hoff, W. D. / Formation of a new buried charge drives a large-amplitude protein quake in photoreceptor activation. In: Biochemistry. 2001 ; Vol. 40, No. 6. pp. 1510-1517.
@article{0215c706b9d546ddb1189b0f2d08b89d,
title = "Formation of a new buried charge drives a large-amplitude protein quake in photoreceptor activation",
abstract = "Photoactive yellow protein (PYP) is a eubacterial photoreceptor and a structural prototype of the PAS domain superfamily of receptor and regulatory proteins. We investigate the activation mechanism of PYP using time-resolved Fourier transform infrared (FTIR) spectroscopy. Our data provide structural, kinetic, and energetic evidence that the putative signaling state of PYP is formed during a large-amplitude protein quake that is driven by the formation of a new buried charge, COO- of the conserved Glu46, in a highly hydrophobic pocket at the active site. A protein quake is a process consisting of global conformational changes that are triggered and driven by a local structural {"}fault{"}. We show that large, global structural changes take place after Glu46 ionization via intramolecular proton transfer to the anionic p-coumarate chromophore, and are suppressed by the absence of COO- formation in the E46Q mutant. Our results demonstrate the significance of buried charge formation in photoreceptor activation. This mechanism may serve as one of the general themes in activation of a range of receptor proteins. In addition, we report the results of time-resolved FTIR spectroscopy of PYP crystals. The direct comparison of time-resolved FTIR spectroscopic data of PYP in aqueous solution and in crystals reveals that the structure of the putative signaling state is not developed in P63 crystals. Therefore, when the structural developments during the functional process of a protein are experimentally determined to be very different in crystals and solutions, one must be cautious in drawing conclusions regarding the functional mechanism of proteins based on time-resolved X-ray crystallography.",
author = "A. Xie and L. Kelemen and J. Hendriks and White, {B. J.} and Hellingwerf, {K. J.} and Hoff, {W. D.}",
year = "2001",
month = "2",
day = "13",
doi = "10.1021/bi002449a",
language = "English",
volume = "40",
pages = "1510--1517",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "6",

}

TY - JOUR

T1 - Formation of a new buried charge drives a large-amplitude protein quake in photoreceptor activation

AU - Xie, A.

AU - Kelemen, L.

AU - Hendriks, J.

AU - White, B. J.

AU - Hellingwerf, K. J.

AU - Hoff, W. D.

PY - 2001/2/13

Y1 - 2001/2/13

N2 - Photoactive yellow protein (PYP) is a eubacterial photoreceptor and a structural prototype of the PAS domain superfamily of receptor and regulatory proteins. We investigate the activation mechanism of PYP using time-resolved Fourier transform infrared (FTIR) spectroscopy. Our data provide structural, kinetic, and energetic evidence that the putative signaling state of PYP is formed during a large-amplitude protein quake that is driven by the formation of a new buried charge, COO- of the conserved Glu46, in a highly hydrophobic pocket at the active site. A protein quake is a process consisting of global conformational changes that are triggered and driven by a local structural "fault". We show that large, global structural changes take place after Glu46 ionization via intramolecular proton transfer to the anionic p-coumarate chromophore, and are suppressed by the absence of COO- formation in the E46Q mutant. Our results demonstrate the significance of buried charge formation in photoreceptor activation. This mechanism may serve as one of the general themes in activation of a range of receptor proteins. In addition, we report the results of time-resolved FTIR spectroscopy of PYP crystals. The direct comparison of time-resolved FTIR spectroscopic data of PYP in aqueous solution and in crystals reveals that the structure of the putative signaling state is not developed in P63 crystals. Therefore, when the structural developments during the functional process of a protein are experimentally determined to be very different in crystals and solutions, one must be cautious in drawing conclusions regarding the functional mechanism of proteins based on time-resolved X-ray crystallography.

AB - Photoactive yellow protein (PYP) is a eubacterial photoreceptor and a structural prototype of the PAS domain superfamily of receptor and regulatory proteins. We investigate the activation mechanism of PYP using time-resolved Fourier transform infrared (FTIR) spectroscopy. Our data provide structural, kinetic, and energetic evidence that the putative signaling state of PYP is formed during a large-amplitude protein quake that is driven by the formation of a new buried charge, COO- of the conserved Glu46, in a highly hydrophobic pocket at the active site. A protein quake is a process consisting of global conformational changes that are triggered and driven by a local structural "fault". We show that large, global structural changes take place after Glu46 ionization via intramolecular proton transfer to the anionic p-coumarate chromophore, and are suppressed by the absence of COO- formation in the E46Q mutant. Our results demonstrate the significance of buried charge formation in photoreceptor activation. This mechanism may serve as one of the general themes in activation of a range of receptor proteins. In addition, we report the results of time-resolved FTIR spectroscopy of PYP crystals. The direct comparison of time-resolved FTIR spectroscopic data of PYP in aqueous solution and in crystals reveals that the structure of the putative signaling state is not developed in P63 crystals. Therefore, when the structural developments during the functional process of a protein are experimentally determined to be very different in crystals and solutions, one must be cautious in drawing conclusions regarding the functional mechanism of proteins based on time-resolved X-ray crystallography.

UR - http://www.scopus.com/inward/record.url?scp=0035852878&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035852878&partnerID=8YFLogxK

U2 - 10.1021/bi002449a

DO - 10.1021/bi002449a

M3 - Article

VL - 40

SP - 1510

EP - 1517

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 6

ER -