ESR STUDY OF DIPHTHAMIDE BIOSYNTHESIS
ESR STUDY OF DIPHTHAMIDE BIOSYNTHESIS
批准号:
8364046
负责人:
BORIS G DZIKOVSKI
金额:
$0.05万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2011
资助国家:
美国
项目状态:
已结题
起止时间:
2011-09-01 至 2012-08-31
关键词:
AnabolismArchaeaBindingBiochemicalBiochemical ReactionElectron Spin Resonance SpectroscopyEnzymesFundingGenesGoalsGrantGuanosine Triphosphate PhosphohydrolasesHumanIronMethodsNational Center for Research ResourcesPeptide Elongation Factor 2PlayPrincipal InvestigatorProtein BiosynthesisProteinsResearchResearch InfrastructureResourcesRoleShapesSourceSpin TrappingSulfurTechnologyTumor Suppressor GenesUnited States National Institutes of HealthWorkYeastsadductcold temperaturecostdimernoveloxidation
中文摘要
这个子项目是利用这些资源的众多研究子项目之一
英文摘要
This subproject is one of many research subprojects utilizing the resources
provided by a Center grant funded by NIH/NCRR. Primary support for the subproject
and the subproject's principal investigator may have been provided by other sources,
including other NIH sources. The Total Cost listed for the subproject likely
represents the estimated amount of Center infrastructure utilized by the subproject,
not direct funding provided by the NCRR grant to the subproject or subproject staff.
The goal or this project is to understand the enzymatic reaction mechanism of diphthamide biosynthesis. Diphthamide is a posttranslationally modified His residue in eukaryotic translation elongation factor 2 (EF-2) a GTPase required for the translocation step of ribosomal protein synthesis. One of the most intriguing facts about diphthamide is that one of the genes involved in its biosynthesis is found to be a tumor suppressor gene in human. Recently, Lin group provided structural and biochemical evidence showing that the first step of diphthamide biosynthesis in archaea uses a novel iron-sulfur cluster enzyme, DPH2 (Zhu and others 2009). ESR spectroscopy at 4- 15K in ACERT played a crucial role in identifying the [4Fe-4S] iron-sulfur cluster.
The results obtained by different methods in Lin's group suggest that unlike known radical SAM enzymes, DPH2 does not form 5¿-deoxyadenosyl radicals. Instead, it breaks the other C-S bond of SAM and transfers the 3-amino-3-carboxylpropyl group to EF-2, possibly via a radical mechanism. To prove or disprove the proposed radical mechanism, we plan to spin-trap and identify the intermediates by the shape of ESR spectra of their adducts with DMPO and possibly other spin-traps. We recently detected some spin trapping adducts and work on their identification. Also, our next step will be a study of other proteins involved in the biosynthesis of diphthamide: yeast DPH1-DPH2 dimer, and yeast DPH3, DPH4. We expect that all these proteins have a Fe-S cluster bound. To identify the cluster, its spin and oxidation state we will record low temperature EPR at a wide field range. It will allow us to detect higher spin states (S= 3/2, 5/2, 7/2) or mixtures thereof.
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会议论文
PRACTICAL ESR COURSE FOR STUDENTS OF INORGANIC CHEMISTRY
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批准号:8364090
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项目类别:
-
资助金额:$0.89万
-
财政年份:2011
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负责人:BORIS G DZIKOVSKI
-
依托单位:
DETECTION OF FREE RADICALS IN TOBACCO SMOKE
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批准号:8364012
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项目类别:
-
资助金额:$0.08万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
INCORPORATION OF SPIN-LABELED ORGANIC MOLECULES INTO CARBON NANOTUBES
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批准号:8364060
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项目类别:
-
资助金额:$0.08万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
ESR STUDY OF BACTERIAL CYTOCHROME C
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批准号:8364045
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项目类别:
-
资助金额:$0.27万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
STRUCTURE OF TRANSITION METAL COMPLEXES BY ESR
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批准号:8364043
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项目类别:
-
资助金额:$1.32万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
CHAPTER FOR NEW VOLUME ON MULTIFREQUENCY EPR
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批准号:8364048
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项目类别:
-
资助金额:$0.05万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
STUDY OF SPIN LABELED WALP PEPTIDES
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批准号:8364047
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项目类别:
-
资助金额:$0.05万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
NON-CHANNEL FORMS OF GRAMICIDIN IN LIPID MEMBRANE BY DQC ESR
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批准号:8363972
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项目类别:
-
资助金额:$0.57万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
ASSESSING NITRIC OXIDE IN PLANT DEFENSE USING NO SPIN TRAPPING
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批准号:8364044
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项目类别:
-
资助金额:$0.05万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
ESR STUDY OF IRON COMPOUNDS FOR UNDERSTANDING NITROGEN FIXATION
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批准号:8364086
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项目类别:
-
资助金额:$0.05万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
GUEST LECTURES IN APPLIED AND ENGINEERING PHYSICS, BIOPHYSICAL METHODS
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批准号:8363961
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项目类别:
-
资助金额:$0.08万
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财政年份:2011
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负责人:BORIS G DZIKOVSKI
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依托单位:
INCLUSION OF SPIN-LABELED MOLECULES IN THE SOLID PHASE OF CYCLODEXTRINS
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批准号:8172136
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项目类别:
-
资助金额:$1.17万
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财政年份:2010
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负责人:BORIS G DZIKOVSKI
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依托单位:
GUEST LECTURES IN APPLIED AND ENGINEERING PHYSICS, BIOPHYSICAL METHODS
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批准号:8172095
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项目类别:
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资助金额:$0.3万
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财政年份:2010
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负责人:BORIS G DZIKOVSKI
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依托单位:
ESR STUDY OF BACTERIAL CYTOCHROME C
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批准号:8172208
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项目类别:
-
资助金额:$0.14万
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财政年份:2010
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负责人:BORIS G DZIKOVSKI
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依托单位:
INCORPORATION OF SPIN-LABELED ORGANIC MOLECULES INTO CARBON NANOTUBES
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批准号:8172225
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项目类别:
-
资助金额:$0.86万
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财政年份:2010
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负责人:BORIS G DZIKOVSKI
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依托单位:
NON-CHANNEL FORMS OF GRAMICIDIN IN LIPID MEMBRANE BY DQC ESR
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批准号:8172109
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项目类别:
-
资助金额:$0.39万
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财政年份:2010
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负责人:BORIS G DZIKOVSKI
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依托单位:
CHAPTER FOR NEW VOLUME OF ?BIOLOGICAL MAGNETIC RESONANCE? SERIES
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批准号:8172211
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项目类别:
-
资助金额:$0.26万
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财政年份:2010
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负责人:BORIS G DZIKOVSKI
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依托单位:
STRUCTURAL EQUATION MODELING OF PIGMENTATION IN DROSOPHILA
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批准号:8172171
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项目类别:
-
资助金额:$0.03万
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财政年份:2010
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负责人:BORIS G DZIKOVSKI
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依托单位:
STRUCTURE OF TRANSITION METAL COMPLEXES BY ESR
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批准号:8172206
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项目类别:
-
资助金额:$0.14万
-
财政年份:2010
-
负责人:BORIS G DZIKOVSKI
-
依托单位:
ASSESSING NITRIC OXIDE IN PLANT DEFENSE USING NO SPIN TRAPPING
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批准号:8172207
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项目类别:
-
资助金额:$0.03万
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财政年份:2010
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负责人:BORIS G DZIKOVSKI
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依托单位:
海外基金