Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein

PloS One
Katrin RöllenAndreas M Stadler

Abstract

Light, oxygen, voltage (LOV) photoreceptors consist of conserved photo-responsive domains in bacteria, archaea, plants and fungi, and detect blue-light via a flavin cofactor. We investigated the blue-light induced conformational transition of the dimeric photoreceptor PpSB1-LOV-R66I from Pseudomonas putida in solution by using small-angle X-ray scattering (SAXS). SAXS experiments of the fully populated light- and dark-states under steady-state conditions revealed significant structural differences between the two states that are in agreement with the known structures determined by crystallography. We followed the transition from the light- to the dark-state by using SAXS measurements in real-time. A two-state model based on the light- and dark-state conformations could describe the measured time-course SAXS data with a relaxation time τREC of ~ 34 to 35 min being larger than the recovery time found with UV/vis spectroscopy. Unlike the flavin chromophore-based UV/vis method that is sensitive to the local chromophore environment in flavoproteins, SAXS-based assay depends on protein conformational changes and provides with an alternative to measure the recovery kinetics.

References

Feb 28, 1998·Current Biology : CB·C P Ponting, L Aravind
Jul 21, 1999·Proceedings of the National Academy of Sciences of the United States of America·J M ChristieW R Briggs
Mar 15, 2001·Proceedings of the National Academy of Sciences of the United States of America·S Crosson, K Moffat
Jul 10, 2001·The Journal of Biological Chemistry·T E SwartzR A Bogomolni
Apr 20, 2002·Biophysical Journal·Aba LosiWolfgang Gärtner
Mar 26, 2003·Biochemistry·John T M KennisRienk van Grondelle
Jul 15, 2003·Photochemistry and Photobiology·Christian Neiss, Peter Saalfrank
Sep 13, 2003·Science·Shannon M HarperKevin H Gardner
Mar 18, 2004·Journal of the American Chemical Society·Shannon M HarperKevin H Gardner
Dec 22, 2004·Biochemistry·Shannon M HarperKevin H Gardner
May 19, 2007·Science·Brian D ZoltowskiBrian R Crane
Aug 22, 2008·Journal of the American Chemical Society·Jessica S LambLois Pollack
Oct 22, 2008·Current Opinion in Plant Biology·Emilie Demarsy, Christian Fankhauser
Aug 29, 2009·Journal of Molecular Biology·Jessica S LambLois Pollack
Mar 3, 2010·Annual Review of Plant Biology·Andreas MöglichKeith Moffat
Jun 19, 2010·Journal of Structural Biology·Haydyn D T Mertens, Dmitri I Svergun
Aug 9, 2011·Nature Reviews. Microbiology·Julien Herrou, Sean Crosson
Dec 6, 2011·Annual Review of Plant Biology·Aba Losi, Wolfgang Gärtner
Feb 14, 2012·Journal of Molecular Biology·Franco CircoloneRenu Batra-Safferling
Apr 17, 2012·Acta Crystallographica. Section D, Biological Crystallography·Willy Wriggers
Apr 27, 2013·Nature·Robert P Rambo, John A Tainer
Jun 15, 2013·Journal of Synchrotron Radiation·Petra PernotSean McSweeney
Dec 10, 2013·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Andrei S Halavaty, Keith Moffat
Apr 1, 2012·Journal of Applied Crystallography·Maxim V PetoukhovDmitri I Svergun
Mar 2, 2016·Proceedings of the National Academy of Sciences of the United States of America·Spencer T GlantzBrian Y Chow
Mar 10, 2016·Biophysical Journal·Andreas M StadlerUlrich Krauss
May 5, 2017·Scientific Reports·Christopher EngelhardRobert Bittl

❮ Previous
Next ❯

Citations

May 23, 2019·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Vera V NazarenkoIvan Gushchin
Feb 24, 2021·The FEBS Journal·Vladimir ArinkinRenu Batra-Safferling
Sep 19, 2020·The Journal of Physical Chemistry Letters·Tayyaba YounasLizhong He

❮ Previous
Next ❯

Methods Mentioned

BETA
X-ray
size exclusion chromatography
Small-angle
NMR
electron-electron double resonance
Size
Dynamic light scattering

Software Mentioned

DAMAVER
CORAL
ATSAS
DAMMIN
SITUS
EOM
DATGNOM

Related Concepts

Related Feeds

Bacterial Protein Structures

Bacterial protein structures can expedite the development of novel antibiotics. Here is the latest research on bacterial proteins and the resolution of their structures.

Bacterial Protein Structures (ASM)

Bacterial protein structures can expedite the development of novel antibiotics. Here is the latest research on bacterial proteins and the resolution of their structures.