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Trans-Membrane Signaling in the Cytochrome b6f Lipoprotein Complex

Award Number: R01GM038323
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 04/01/1987
PERIOD OF PERFORMANCE END DATE: 12/31/2018

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Trans-Membrane Signaling in the Cytochrome b6f Lipoprotein Complex - DESCRIPTION (provided by applicant): (1) The system. The cytochrome b6f complex is one of only 60 independent hetero-oligomeric membrane proteins in the data bank. (2) Recent achievements, structure-function: (a) a newly recognized unique heme in the complex was characterized; (b) the quinone dependent pathway for proton translocation was described at a resolution of 2.7 A (c) a unique function of the bound chlorophyll in occluding the quinone entry portal was demonstrated; (d) detailed lipid analysis was undertaken, conservation shown of many lipid functions between the b6f complex and the mitochondrial cytochrome bc1 complex, and (e) conformational changes shown to be induced by anionic lipid. (3) Proposed studies: (a) Using His-tagged cytochrome b6f complex derived from C. reinhardtii, (b) crystal structures of higher order ("super")-complexes of the electron transport complexes in the photosynthetic electron transport chain will be sought, following the precedent set for such super-complexes in the respiratory chain. A starting point is utilization of an isolated complex of the 230 kDa cytochrome b6f complex with the 1 MDa trimeric photosystem I reaction center. Elucidation of the structure of such a complex would likely describe new pathways for electronic communication between the complexes via quinones and soluble electron transport proteins, and be used to define the interactions and functions of the lipids internal to the cytochrome complex. (c) The stoichiometry and arrangement of the lipids, and their effects on the dynamics of the cytochrome complex, will be analyzed by native and hydrogen-deuterium mass spectroscopy. (d) Assembly; role of lipids. To test the role of the lipids in separating different domains of the cytochrome complex, mass spectroscopic analysis, particularly native MS, will be applied to analysis of lipid interactions at different stages of assembly of the b6f complex. (e Dielectric heterogeneity of the b6f complex; dependence of structure on electric field. Our studies imply that the internal dielectric constant within the b6f complex is heterogeneous, i. e., different between the different heme pairs within the complex. Virtually all membrane proteins, certainly those involved in energy-transduction and active transport, function in the presence of large electrical fields. With the long-term goal of determining the effect of electric fields on membrane protein structures, the effect of such fields on cytochrome spectra, redox properties, and secondary structure assayed by far-UV circular dichroism spectra, will be studied. (f) Trans- membrane signaling; structure of the signaling complex. The redox signaling system in the cytochrome b6f complex uses a unique trans-membrane signaling mechanism dependent upon oxidation of plastoquinol on the electro-positive side of the complex. A crystal structure will be sought of the trans-membrane signaling complex consisting of the b6f complex reconstituted with the protein kinase. These studies are directed not only to better crystallographic resolution and understanding of the structure of the cytochrome complex as a hetero- oligomeric membrane lipoprotein that is relevant to its function of trans-membrane signaling.


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: 2019 ( Subtotal = -$1,183 )
2019	2017	PURDUE UNIVERSITY	155 S GRANT ST	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	28	5/7/2019	NON-COMPETING CONTINUATION	-$1,183
														Subtotal = -$1,183

Issue Date FY: 2017 ( Subtotal = $405,262 )
2017	2017	PURDUE UNIVERSITY	401 SOUTH GRANT ST	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	28	12/15/2016	NON-COMPETING CONTINUATION	$364,735
2017	2017	PURDUE UNIVERSITY	401 SOUTH GRANT ST	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	001	28	6/5/2017	NON-COMPETING CONTINUATION	$40,527
														Subtotal = $405,262

Issue Date FY: 2016 ( Subtotal = $498,649 )
2016	2016	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	001	27	3/18/2016	NON-COMPETING CONTINUATION	$34,650
2016	2016	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	002	27	9/8/2016	SUPPLEMENT FOR EXPANSION	$152,149
2016	2016	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	27	1/21/2016	NON-COMPETING CONTINUATION	$311,850
														Subtotal = $498,649

Issue Date FY: 2015 ( Subtotal = $346,500 )
2015	2015	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	001	26	3/16/2015	NON-COMPETING CONTINUATION	$34,650
2015	2015	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	26	12/8/2014	NON-COMPETING CONTINUATION	$311,850
														Subtotal = $346,500

Issue Date FY: 2014 ( Subtotal = $343,615 )
2014	2014	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	001	25	12/20/2013	COMPETING CONTINUATION	$343,615
2014	2012	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	24	12/17/2013	NON-COMPETING CONTINUATION	$0
														Subtotal = $343,615

Issue Date FY: 2012 ( Subtotal = $332,442 )
2012	2012	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	001	24	3/7/2012	NON-COMPETING CONTINUATION	$33,244
2012	2012	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	24	1/23/2012	NON-COMPETING CONTINUATION	$299,198
														Subtotal = $332,442

Issue Date FY: 2011 ( Subtotal = $329,905 )
2011	2011	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	001	23	6/6/2011	NON-COMPETING CONTINUATION	$29,992
2011	2011	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	23	2/8/2011	NON-COMPETING CONTINUATION	$299,913
														Subtotal = $329,905

Issue Date FY: 2010 ( Subtotal = $330,659 )
2010	2010	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	22	1/27/2010	NON-COMPETING CONTINUATION	$330,659
														Subtotal = $330,659

Issue Date FY: 2009 ( Subtotal = $331,487 )
2009	2009	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	21	3/17/2009	COMPETING CONTINUATION	$331,487
														Subtotal = $331,487

Issue Date FY: 2006 ( Subtotal = $312,394 )
2006	2006	PURDUE UNIVERSITY	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	20	6/27/2006	NON-COMPETING CONTINUATION	$312,394
														Subtotal = $312,394

Issue Date FY: 2005 ( Subtotal = $316,909 )
2005	2005	PURDUE UNIVERSITY	SPONS PROGRAMS-1063 HOVDE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	19	7/11/2005	NON-COMPETING CONTINUATION	$316,909
														Subtotal = $316,909

Issue Date FY: 2004 ( Subtotal = $315,189 )
2004	2004	PURDUE UNIVERSITY	SPONS PROGRAMS-1063 HOVDE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	18	6/21/2004	NON-COMPETING CONTINUATION	$315,189
														Subtotal = $315,189

Issue Date FY: 2003 ( Subtotal = $462,471 )
2003	2003	PURDUE RESEARCH FOUNDATION	SPONS PROGRAMS-1063 HOVDE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	16		NON-COMPETING CONTINUATION	$147,700
2003	2003	PURDUE UNIVERSITY	SPONS PROGRAMS-1063 HOVDE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	17	6/9/2003	COMPETING CONTINUATION	$314,771
														Subtotal = $462,471

Issue Date FY: 2002 ( Subtotal = $252,164 )
2002	2002	PURDUE RESEARCH FOUNDATION	SPONS PROGRAMS-1063 HOVDE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	15	11/21/2001	NON-COMPETING CONTINUATION	$252,164
														Subtotal = $252,164

Issue Date FY: 2001 ( Subtotal = $245,108 )
2001	2001	PURDUE RESEARCH FOUNDATION	SPONS PROGRAMS-1063 HOVDE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	14	1/8/2001	NON-COMPETING CONTINUATION	$245,108
														Subtotal = $245,108

Issue Date FY: 2000 ( Subtotal = $240,244 )
2000	2000	PURDUE RESEARCH FOUNDATION	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	13	12/22/1999	NON-COMPETING CONTINUATION	$240,244
														Subtotal = $240,244

Issue Date FY: 1999 ( Subtotal = $173,435 )
1999	1999	PURDUE RESEARCH FOUNDATION	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	12	7/26/1999	COMPETING CONTINUATION	$100,739
1999	1999	PURDUE RESEARCH FOUNDATION	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	001	11	12/24/1998	SUPPLEMENT FOR EXPANSION	$72,696
														Subtotal = $173,435

Issue Date FY: 1998 ( Subtotal = $216,909 )
1998	1998	PURDUE RESEARCH FOUNDATION	610 PURDUE HALL	WEST LAFAYETTE	IN	47907	TIPPECANOE	USA	Biomedical Research and Research Training	000	11	11/28/1997	NON-COMPETING CONTINUATION	$216,909
														Subtotal = $216,909

Grand Total All Awards = $5,452,159

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Trans-Membrane Signaling in the Cytochrome b6f Lipoprotein Complex

Award Number: R01GM038323

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 04/01/1987

PERIOD OF PERFORMANCE END DATE: 12/31/2018

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