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Taking Snapshots of the Water-Oxidation Reaction by the Mn4Ca Oxygen-Evolving Complex Using X-ray Crystallography and Spectroscopy

Award Number: R01GM055302
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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Project Summary/Abstract Metalloproteins containing manganese in a redox-active role are involved in a variety of physiologically important reactions of dioxygen metabolism. Perhaps the most complex is the Mn4CaO5 cluster that is involved in the oxidation of water to dioxygen in photosystem II (PS II), an ~500 kDa multi-subunit membrane protein complex. The water-oxidation reaction in PS II involves removal of four electrons from two water molecules, in a stepwise manner by light-induced oxidation, to produce a molecule of oxygen. PS II and the Mn4CaO5 cluster generate almost all of the dioxygen that supports aerobic life, and it is abundant in the atmosphere because of its constant regeneration by the oxidation of water. The light-induced oxidation of water to dioxygen is one of the most important chemical processes occurring on such a large scale in the biosphere. Although the structure of PS II and the chemistry at the catalytic site have been studied intensively, understanding the sequence in the chemistry at atomic-scale from light absorption to water-oxidation requires a new approach beyond the conventional steady state X-ray crystallography and X-ray spectroscopy at cryogenic temperatures. Following the dynamic changes in the structure of PS II and the Mn4CaO5 cluster at ambient conditions at physiological temperatures, while overcoming the severe X-ray damage to the redox active center is key for deriving the mechanism. The intense and ultra-short femtosecond (fs) X-ray pulses from a X-ray free electron laser (XFEL) provide an opportunity to overcome the current limitations in room temperature data collection for biological samples at traditional X-ray sources. The fs X-ray pulses allow us to acquire the signal before the sample is destroyed, thus making the light-induced snapshot study proposed here possible. The objective of this proposal is to study the protein structure and dynamics of PS II with X-ray diffraction, as well as the chemical structure and changes in the Mn4CaO5 cluster (charge and spin density, and covalency) with X-ray spectroscopy during the light-driven process of PS II. We will use the XFEL facilities at Stanford and elsewhere to collect X-ray diffraction and emission spectra simultaneously, and X-ray absorption spectra of the Mn cluster in its native and intermediates states at room temperature in a time-resolved manner, to capture short-lived intermediates and the step that includes the O-O bond formation. We have also started studying the process of assembly of the Mn cluster as repair and assembly of PS II is an essential component in nature. These studies have the potential to provide an unprecedented combination of correlated data between the PS II protein, the co-factors, and the Mn4CaO5 cluster, providing the geometric and electronic structure and the changes that occur during the catalytic cycle, all of which are necessary for a complete understanding of the mechanism of water oxidation. !


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2022 ( Subtotal = $646,351 )
2022	2022	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	000	24	5/4/2022	NON-COMPETING CONTINUATION	$646,351
														Subtotal = $646,351

Issue Date FY: 2021 ( Subtotal = $568,314 )
2021	2021	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	000	23	4/8/2021	NON-COMPETING CONTINUATION	$568,314
														Subtotal = $568,314

Issue Date FY: 2020 ( Subtotal = $566,500 )
2020	2020	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	002	22	4/24/2020	NON-COMPETING CONTINUATION	$566,500
2020	2019	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	001	21	2/3/2020	COMPETING CONTINUATION	$183,097
2020	2018	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	000	20	2/3/2020	NON-COMPETING CONTINUATION	-$183,097
														Subtotal = $566,500

Issue Date FY: 2019 ( Subtotal = $626,027 )
2019	2019	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	001	21	4/25/2019	COMPETING CONTINUATION	$569,056
2019	2019	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	002	21	8/13/2019	SUPPLEMENT FOR EXPANSION	$56,971
2019	2018	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	000	20	4/22/2019	NON-COMPETING CONTINUATION	$0
														Subtotal = $626,027

Issue Date FY: 2018 ( Subtotal = $531,690 )
2018	2018	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	001	20	5/21/2018	NON-COMPETING CONTINUATION	$53,169
2018	2018	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	000	20	1/5/2018	NON-COMPETING CONTINUATION	$478,521
														Subtotal = $531,690

Issue Date FY: 2017 ( Subtotal = $534,510 )
2017	2017	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	000	19	1/5/2017	NON-COMPETING CONTINUATION	$481,061
2017	2017	REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE	1 CYCLOTRON RD	BERKELEY	CA	94720	ALAMEDA	USA	Biomedical Research and Research Training	001	19	6/5/2017	NON-COMPETING CONTINUATION	$53,449
														Subtotal = $534,510

Issue Date FY: 2016 ( Subtotal = $528,270 )
2016	2016	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	18	3/8/2016	NON-COMPETING CONTINUATION	$528,270
														Subtotal = $528,270

Issue Date FY: 2015 ( Subtotal = $520,690 )
2015	2015	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	002	16	8/27/2015	SUPPLEMENT FOR EXPANSION	$0
2015	2015	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	16	1/29/2015	SUPPLEMENT FOR EXPANSION	$164,162
2015	2015	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	004	17	9/1/2015	COMPETING CONTINUATION	$356,528
2015	2013	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	001	16	1/29/2015	NON-COMPETING CONTINUATION	$0
2015	2013	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	003	16	8/27/2015	NON-COMPETING CONTINUATION	$0
														Subtotal = $520,690

Issue Date FY: 2013 ( Subtotal = $464,404 )
2013	2013	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	16	8/13/2013	NON-COMPETING CONTINUATION	$464,404
														Subtotal = $464,404

Issue Date FY: 2012 ( Subtotal = $481,000 )
2012	2012	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	15	8/13/2012	NON-COMPETING CONTINUATION	$481,000
														Subtotal = $481,000

Issue Date FY: 2011 ( Subtotal = $502,924 )
2011	2011	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	14	9/15/2011	NON-COMPETING CONTINUATION	$502,924
														Subtotal = $502,924

Issue Date FY: 2010 ( Subtotal = $501,949 )
2010	2010	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	13	9/1/2010	COMPETING CONTINUATION	$501,949
														Subtotal = $501,949

Issue Date FY: 2009 ( Subtotal = $0 )
2009	2008	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	12	9/9/2009	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2008 ( Subtotal = $315,841 )
2008	2008	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	12	3/12/2008	NON-COMPETING CONTINUATION	$315,841
														Subtotal = $315,841

Issue Date FY: 2007 ( Subtotal = $316,181 )
2007	2007	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	11	3/16/2007	NON-COMPETING CONTINUATION	$316,181
														Subtotal = $316,181

Issue Date FY: 2006 ( Subtotal = $316,531 )
2006	2006	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	10	3/31/2006	NON-COMPETING CONTINUATION	$316,531
														Subtotal = $316,531

Issue Date FY: 2005 ( Subtotal = $315,104 )
2005	2005	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	9	3/24/2005	COMPETING CONTINUATION	$315,104
														Subtotal = $315,104

Issue Date FY: 2004 ( Subtotal = $346,192 )
2004	2004	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	001	8	9/27/2004	SUPPLEMENT FOR EXPANSION	$102,992
2004	2004	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	8	12/17/2003	NON-COMPETING CONTINUATION	$243,200
														Subtotal = $346,192

Issue Date FY: 2003 ( Subtotal = $232,028 )
2003	2003	LAWRENCE BERKELEY LABORATORY	POST OFFICE BOX 528	BERKELEY	CA	94701	ALAMEDA	USA	Biomedical Research and Research Training	000	7		NON-COMPETING CONTINUATION	$232,028
														Subtotal = $232,028

Issue Date FY: 2002 ( Subtotal = $229,701 ) (Continued on the next page)
Grand Total All Awards = $9,535,056

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Taking Snapshots of the Water-Oxidation Reaction by the Mn4Ca Oxygen-Evolving Complex Using X-ray Crystallography and Spectroscopy

Award Number: R01GM055302

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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