skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Binding of red form of Orange Carotenoid Protein (OCP) to phycobilisome is not sufficient for quenching

Journal Article · · Biochimica et Biophysica Acta - Bioenergetics

The Orange Carotenoid Protein (OCP) is responsible for photoprotection in many cyanobacteria. Absorption of blue light drives the conversion of the orange, inactive form (OCPO) to the red, active form (OCPR). Concomitantly, the N–terminal domain (NTD) and the C–terminal domain (CTD) of OCP separate, which ultimately leads to the formation of a quenched OCPR–PBS complex. The details of the photoactivation of OCP have been intensely researched. Binding site(s) of OCPR on the PBS core have also been proposed. Yet, the post–binding events of the OCPR–PBS complex remain unclear. In this work, we demonstrate that PBS–bound OCPR is not sufficient as a PBS excitation energy quencher. Using site–directed mutagenesis, we generated a suite of single point mutations at OCP Leucine 51 (L51) of Synechocystis 6803. Steady–state and time–resolved fluorescence analyses demonstrated that all mutant proteins are unable to quench the PBS fluorescence, owing to either failed OCP binding to PBS, or, if bound, an OCP–PBS quenching state failed to form. The SDS–PAGE and Western blot analysis support that the L51A (Alanine) mutant binds to the PBS and therefore belongs to the second category. We hypothesize that upon binding to PBS, OCPR likely reorganizes and adopts a new conformational state (OCP3rd) different than either OCPO or OCPR to allow energy quenching, depending on the cross–talk between OCPR and its PBS core–binding counterpart.

Research Organization:
Washington Univ., St. Louis, MO (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences, and Biosciences Division
Grant/Contract Number:
FG02-07ER15902; SC0001035; DE–FG02–07ER15902; DE–SC0001035
OSTI ID:
1768117
Alternate ID(s):
OSTI ID: 1593978; OSTI ID: 1594183
Journal Information:
Biochimica et Biophysica Acta - Bioenergetics, Journal Name: Biochimica et Biophysica Acta - Bioenergetics Vol. 1861 Journal Issue: 3; ISSN 0005-2728
Publisher:
ElsevierCopyright Statement
Country of Publication:
Netherlands
Language:
English
Citation Metrics:
Cited by: 5 works
Citation information provided by
Web of Science

References (45)

Phycobilisome-reaction centre interaction in cyanobacteria journal October 2007
Carotenoid–protein interaction alters the S1 energy of hydroxyechinenone in the Orange Carotenoid Protein journal March 2013
Excited State Properties of 3′-Hydroxyechinenone in Solvents and in the Orange Carotenoid Protein from Synechocystis sp. PCC 6803 journal May 2014
The Orange Carotenoid Protein: a blue-green light photoactive protein journal January 2013
A Molecular Mechanism for Nonphotochemical Quenching in Cyanobacteria journal May 2017
A carotenoid-protein from cyanobacteria journal October 1981
A 12 A carotenoid translocation in a photoswitch associated with cyanobacterial photoprotection journal June 2015
The Photophysics of the Orange Carotenoid Protein, a Light-Powered Molecular Switch journal February 2012
Site, Rate, and Mechanism of Photoprotective Quenching in Cyanobacteria journal November 2011
Hybrid coupling of R-phycoerythrin and the orange carotenoid protein supports the FRET-based mechanism of cyanobacterial photoprotection journal August 2019
The Signaling State of Orange Carotenoid Protein journal August 2015
Phycobilisome rod mutants in Synechocystis sp. strain PCC6803 journal December 2004
Site of non-photochemical quenching of the phycobilisome by orange carotenoid protein in the cyanobacterium Synechocystis sp. PCC 6803 journal August 2012
Evolution of flexible non-photochemical quenching mechanisms that regulate light harvesting in oxygenic photosynthesis journal June 2013
Investigation of OCP-triggered dissipation of excitation energy in PSI/PSII-less Synechocystis sp. PCC 6803 mutant using non-linear laser fluorimetry journal July 2012
Identification of a mechanism of photoprotective energy dissipation in higher plants journal November 2007
The Essential Role of the N-Terminal Domain of the Orange Carotenoid Protein in Cyanobacterial Photoprotection: Importance of a Positive Charge for Phycobilisome Binding journal May 2012
A photoactive carotenoid protein acting as light intensity sensor journal August 2008
Specificity of the Cyanobacterial Orange Carotenoid Protein: Influences of Orange Carotenoid Protein and Phycobilisome Structures journal December 2013
Role of inter-domain cavity in the attachment of the orange carotenoid protein to the phycobilisome core and to the fluorescence recovery protein journal July 2015
Cu + Contributes to the Orange Carotenoid Protein-Related Phycobilisome Fluorescence Quenching and Photoprotection in Cyanobacteria journal June 2019
A Soluble Carotenoid Protein Involved in Phycobilisome-Related Energy Dissipation in Cyanobacteria journal March 2006
Picosecond Kinetics of Light Harvesting and Photoprotective Quenching in Wild-Type and Mutant Phycobilisomes Isolated from the Cyanobacterium Synechocystis PCC 6803 journal April 2012
The ConSurf-DB: pre-calculated evolutionary conservation profiles of protein structures journal October 2008
Correction to: Site, trigger, quenching mechanism and recovery of non-photochemical quenching in cyanobacteria: recent updates journal April 2018
Mass spectrometry footprinting reveals the structural rearrangements of cyanobacterial orange carotenoid protein upon light activation journal December 2014
Photoactivation and relaxation studies on the cyanobacterial orange carotenoid protein in the presence of copper ion journal March 2017
ConSurf 2005: the projection of evolutionary conservation scores of residues on protein structures journal July 2005
Structural and Functional Modularity of the Orange Carotenoid Protein: Distinct Roles for the N- and C-Terminal Domains in Cyanobacterial Photoprotection journal January 2014
The photocycle of orange carotenoid protein conceals distinct intermediates and asynchronous changes in the carotenoid and protein components journal November 2017
Orange carotenoid protein burrows into the phycobilisome to provide photoprotection journal March 2016
Excitation energy transfer between photosystem II and photosystem I in red algae: Larger amounts of phycobilisome enhance spillover journal July 2011
In Vitro Reconstitution of the Cyanobacterial Photoprotective Mechanism Mediated by the Orange Carotenoid Protein in Synechocystis PCC 6803 journal July 2011
Photosystem II fluorescence quenching in the cyanobacterium Synechocystis PCC 6803: involvement of two different mechanisms journal April 2000
The Crystal Structure of a Cyanobacterial Water-Soluble Carotenoid Binding Protein journal January 2003
Spectroscopic Properties of the Carotenoid 3‘-Hydroxyechinenone in the Orange Carotenoid Protein from the Cyanobacterium Arthrospira maxima journal March 2005
Cyanobacterial photoprotection by the orange carotenoid protein journal December 2016
Photoprotection in cyanobacteria: the orange carotenoid protein (OCP)-related non-photochemical-quenching mechanism journal May 2007
Biosynthesis of soluble carotenoid holoproteins in Escherichia coli journal March 2015
Molecular Mechanism of Photoactivation and Structural Location of the Cyanobacterial Orange Carotenoid Protein journal December 2013
Regulation of Orange Carotenoid Protein Activity in Cyanobacterial Photoprotection journal July 2015
Phycobilisomes Supply Excitations to Both Photosystems in a Megacomplex in Cyanobacteria journal November 2013
Single-molecule trapping and spectroscopy reveals photophysical heterogeneity of phycobilisomes quenched by Orange Carotenoid Protein journal March 2019
Fluorescence quenching of the phycobilisome terminal emitter LCM from the cyanobacterium Synechocystis sp. PCC 6803 detected in vivo and in vitro journal August 2013
Photoactivation Mechanism, Timing of Protein Secondary Structure Dynamics and Carotenoid Translocation in the Orange Carotenoid Protein journal November 2018