Journal Article

PreJuSER-20004

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Structural basis for the slow dark recovery of a full-length LOV protein from Pseudomonas putida

Circolone, F. ; Granzin, J.FZJ* ; Jentzsch, K. ; Drepper, T.FZJ* ; Jaeger, K. E. ; Willbold, D.FZJ* ; Krauss, U. ; Batra-Safferling, R.FZJ*

2012
Elsevier Amsterdam [u.a.]

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Please use a persistent id in citations: doi:10.1016/j.jmb.2012.01.056

Abstract: Blue-light photoreceptors containing light–oxygen–voltage (LOV) domains regulate a myriad of different physiological responses in both eukaryotes and prokaryotes. Their light sensitivity is intricately linked to the photochemistry of the non-covalently bound flavin mononucleotide (FMN) chromophore that forms a covalent adduct with a conserved cysteine residue in the LOV domain upon illumination with blue light. All LOV domains undergo the same primary photochemistry leading to adduct formation; however, considerable variation is found in the lifetime of the adduct state that varies from seconds to several hours. The molecular mechanism underlying this variation among the structurally conserved LOV protein family is not well understood. Here, we describe the structural characterization of PpSB1-LOV, a very slow cycling full-length LOV protein from the Gram-negative bacterium Pseudomonas putida KT2440. Its crystal structure reveals a novel dimer interface that is mediated by N- and C-terminal auxiliary structural elements and a unique cluster of four arginine residues coordinating with the FMN-phosphate moiety. Site-directed mutagenesis of two arginines (R61 and R66) in PpSB1-LOV resulted in acceleration of the dark recovery reaction approximately by a factor of 280. The presented structural and biochemical data suggest a direct link between structural features and the slow dark recovery observed for PpSB1-LOV. The overall structural arrangement of PpSB1-LOV, together with a complementary phylogenetic analysis, highlights a common ancestry of bacterial LOV photoreceptors and Per-ARNT-Sim chemosensors.

Keyword(s): Bacterial Proteins: chemistry (MeSH) ; Crystallography, X-Ray (MeSH) ; Flavin Mononucleotide: chemistry (MeSH) ; Light (MeSH) ; Photoreceptors, Microbial: chemistry (MeSH) ; Protein Structure, Tertiary (MeSH) ; Pseudomonas putida: metabolism (MeSH) ; Bacterial Proteins ; Photoreceptors, Microbial ; Flavin Mononucleotide ; J ; photoreceptor (auto) ; LOV domain (auto) ; PAS domain (auto) ; crystal structure (auto) ; photocycle (auto)

Note: The authors wish to thank Georg Buldt for continuous generous support and the scientists at beamline ID23-2 at the ESRF (Grenoble, France). Part of this work has been funded by the Deutsche Forschungsgemeinschaft within the GRK1166 "Biocatalysis in Non-Conventional Media".

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Contributing Institute(s):

1. Strukturbiochemie (ICS-6)

Research Program(s):

1. Funktion und Dysfunktion des Nervensystems (P33)
2. BioSoft: Makromolekulare Systeme und biologische Informationsverarbeitung (P45)

Appears in the scientific report 2012

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Database coverage:
; BIOSIS Previews ; Current Contents - Life Sciences ; JCR ; NCBI Molecular Biology Database ; Nationallizenz ; SCOPUS ; Science Citation Index ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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