Cross-Linking of Transmembrane Helices Reveals a Rigid-Body Mechanism in Bacteriorhodopsin Transport

A flashy protein: The flash‐induced transient protonation of pyranine has been studied in the presence of the cross‐linked double mutant E166C/A228C (red trace), a reduced mutant (blue trace), and wild‐type bacteriorhodopsin (gray traces). If helices F and G are cross‐linked, there is a delay in bot...

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Published inAngewandte Chemie (International ed.) Vol. 48; no. 45; pp. 8523 - 8525
Main Authors Simón-Vázquez, Rosana, Lazarova, Tzvetana, Perálvarez-Marín, Alex, Bourdelande, José-Luis, Padrós, Esteve
Format Journal Article
LanguageEnglish
Published Weinheim WILEY-VCH Verlag 26.10.2009
WILEY‐VCH Verlag
Subjects
Bacteria
Bacterial Proteins - chemistry
Bacterial Proteins - metabolism
Bacteriorhodopsin
Bacteriorhodopsins - chemistry
Bacteriorhodopsins - metabolism
Cross-Linking Reagents - chemistry
Cross-Linking Reagents - pharmacology
Crosslinking
Delay
Flash photolysis
helical structures
Helices
Kinetics
membrane proteins
Membrane Proteins - chemistry
Membrane Proteins - metabolism
Membrane Transport Proteins - chemistry
Protein Conformation
Proteins
proton transport
structural biology
Transport
Uptakes
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Summary:A flashy protein: The flash‐induced transient protonation of pyranine has been studied in the presence of the cross‐linked double mutant E166C/A228C (red trace), a reduced mutant (blue trace), and wild‐type bacteriorhodopsin (gray traces). If helices F and G are cross‐linked, there is a delay in both proton release (extracellular side) and proton uptake (cytoplasmic side). Together with flash photolysis and FTIR studies, these data support a rigid‐body mechanism of bacteriorhodopsin proton transport.
Bibliography:ark:/67375/WNG-C2ZRRNT2-6
ArticleID:ANIE200904031
We thank Dr. Víctor Lórenz-Fonfría and Dr. Pere Garriga for helpful discussions, Dr. Enric Querol for advice at the beginning of this work, Jordi Hernando for help in the flash photolysis experiments, Elodia Serrano and Neus Ontiveros for excellent technical assistance, and Dr. Mathias Lübben (Ruhr-Universität Bochum) for providing the pEF191 vector and the MPK409 strain. This work was supported by a Direcció General de Recerca fellowship (2007PIV10038) to T.L. and by a Dirección General de Investigación grant (BFU2006-04656/BMC).
istex:CEE40F9F81FA463BC656F80189380736041BFF3B
Dirección General de Investigación - No. BFU2006-04656/BMC
We thank Dr. Víctor Lórenz‐Fonfría and Dr. Pere Garriga for helpful discussions, Dr. Enric Querol for advice at the beginning of this work, Jordi Hernando for help in the flash photolysis experiments, Elodia Serrano and Neus Ontiveros for excellent technical assistance, and Dr. Mathias Lübben (Ruhr‐Universität Bochum) for providing the pEF191 vector and the MPK409 strain. This work was supported by a Direcció General de Recerca fellowship (2007PIV10038) to T.L. and by a Dirección General de Investigación grant (BFU2006‐04656/BMC).
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.200904031