Assembly of light-harvesting chlorophyll a/b complex in vitro. Time-resolved fluorescence measurements

Biochemistry
Paula J Booth, H Paulsen

Abstract

The assembly kinetics of a pigment-binding membrane protein, the light- harvesting chlorophyll a/b complex of green plants, have been determined in vitro. Time-resolved fluorescence spectroscopy, with millisecond time resolution, has been used to monitor changes in both protein and chlorophyll (Chl) fluorescence, as well as in energy transfer from Chl b to Chl a, during complex assembly. Three reaction steps could be resolved after rapid, stopped-flow, mixing of the apoprotein (light-harvesting Chl a/b protein, LHCP) and pigments, solubilized in sodium dodecyl sulfate (SDS) and octyl glucoside (OG), respectively. A fast step in the range of 10 ms was detected regardless of whether the reaction mixture contained pigments or proteins, or both, and is interpreted as being connected with the formation of mixed SDS and OG detergent micelles. Two further steps were resolved: one with a time constant of about 1 min and another, slow step with a time constant of several minutes. Both of these were dependent on the presence of protein, Chls, and xanthophylls. Most, if not all, of the energy transfer from Chl b to Chl a was established during the slow step, indicating the the juxtaposition of these pigments, either by a structural rearra...Continue Reading

References

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Citations

Jun 14, 2005·Journal of Molecular Biology·Alexander ProdöhlDirk Schneider
Jun 8, 2002·Journal of Molecular Biology·Ruth Horn, Harald Paulsen
Feb 1, 1997·Current Opinion in Biotechnology·M P Brown, C Royer
Oct 17, 2009·Proceedings of the National Academy of Sciences of the United States of America·Christoph DockterHarald Paulsen
May 26, 2010·The Journal of Biological Chemistry·Sebastian Falk, Irmgard Sinning
Feb 25, 2011·The Journal of Biological Chemistry·Natalie D Di BartoloPaula J Booth
Jul 20, 1999·Annual Review of Biophysics and Biomolecular Structure·U HauptsD Oesterhelt
Apr 6, 2012·Current Opinion in Structural Biology·Paula J Booth
Jul 25, 2006·Biochimica Et Biophysica Acta·Thomas VolkmerDirk Schneider
Nov 17, 2004·Journal of Molecular Biology·Mark Lorch, Paula J Booth
Sep 8, 2004·Journal of Molecular Biology·Samantha J AllenPaula J Booth
Sep 8, 2004·Journal of Molecular Biology·Samantha J AllenPaula J Booth
Mar 31, 2009·Biochimica Et Biophysica Acta·Tiago Barros, Werner Kühlbrandt
Nov 3, 2004·Biochimica Et Biophysica Acta·Annela M SeddonPaula J Booth
Mar 31, 2016·Biophysical Journal·Rashmi PanigrahiM Joanne Lemieux
Jul 4, 2006·Current Opinion in Structural Biology·Paula J Booth, Paul Curnow
Jan 24, 2006·Journal of Molecular Biology·Emma L R ComptonPaula J Booth
Dec 24, 2013·Biochimica Et Biophysica Acta·Mara WerwieHarald Paulsen
Nov 1, 2007·Archives of Biochemistry and Biophysics·Ann Marie Stanley, Karen G Fleming
Nov 5, 2014·Langmuir : the ACS Journal of Surfaces and Colloids·Sebastian RoederHarald Paulsen
Mar 3, 2017·Biochemistry·Daniel M PalmHarald Paulsen
Jun 19, 2004·Molecular Membrane Biology·Paula J Booth, Stephen High
Mar 10, 2005·The Journal of Biological Chemistry·Gunnar JeschkeHarald Paulsen
Jun 29, 2006·The Journal of Biological Chemistry·Stephan HobeHarald Paulsen
Feb 9, 2011·Journal of Molecular Modeling·Andrzej SikorskiSzymon Zerko
Aug 12, 2004·The Journal of Biological Chemistry·Ruth Horn, Harald Paulsen
Apr 28, 2020·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Philipp GirrHarald Paulsen
Oct 10, 2018·Nature Plants·Daniel M PalmHarald Paulsen
Dec 5, 2019·Scientific Reports·Alessandro AgostiniElisabetta Collini
Jan 27, 2019·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·Govind ReddyLingamallu Giribabu
Mar 10, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Mara WerwieHarald Paulsen
Jan 24, 2018·The Journal of Physical Chemistry Letters·Alessandro AgostiniDonatella Carbonera
Sep 11, 2019·The Journal of Physical Chemistry. B·Alessandro AgostiniDonatella Carbonera

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