Defining the Molecular Architecture for Transmembrane Acylation by a Membrane Bound O-Acyltransferase
Defining the Molecular Architecture for Transmembrane Acylation by a Membrane Bound O-Acyltransferase
批准号:
10246913
负责人:
John Daniel Chisholm
金额:
$33.0万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2019
资助国家:
美国
项目状态:
已结题
起止时间:
2019-09-01 至 2024-08-31
关键词:
Active SitesAcylationAcyltransferaseAddressArchitectureBinding SitesBiochemicalBioinformaticsBiologicalCatalytic DomainChemicalsComputer ModelsCoupledCysteineDesire for foodDevelopmentDiabetes MellitusDiseaseDockingEnzyme InhibitionEnzymesErinaceidaeFamilyFamily memberFeeding behaviorsFosteringFoundationsGoalsHomeostasisHumanIntegral Membrane ProteinLocationMalignant NeoplasmsMembraneMembrane ProteinsMetabolismMissionModelingMolecularMutagenesisPathway interactionsPharmacologic SubstancePlayPorcupinesProtein FamilyProteinsPublic HealthReportingResearchResistanceRoleSerineSignal PathwaySignal TransductionStructural ModelsStructureSystemTherapeuticUnited States National Institutes of HealthValidationWorkbasebiochemical toolscancer therapycomputerized toolsdesignghrelinglucose metabolismhuman diseaseinhibitor/antagonistinnovationinsightmembermolecular dynamicsnanomolarnovelpeptide hormonepredictive testprotein acyltransferaseprotein structuresmall moleculetherapeutic targettool
中文摘要
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英文摘要
Acylated secreted proteins play essential roles in intercellular and organismal signaling pathways implicated in
multiple diseases including diabetes and cancer. Protein-modifying members of the membrane-bound O-
acyltransferase (MBOAT) enzyme family constitute key molecular control points for signaling through their roles
in modifying the secreted proteins ghrelin, Hedgehog, and Wnt. Of these, ghrelin is unique in that it regulates
feeding behavior and energy homeostasis. Since ghrelin requires acylation by GOAT for biological activity, a
detailed understanding of this MBOAT family member is imperative to understand the role of ghrelin in disease
and to target ghrelin-dependent pathways. However, the dearth of information regarding the structure, substrate
binding sites, and catalytic mechanism of GOAT impedes understanding ghrelin signaling and development of
small molecule tools to study the role of GOAT in metabolism-related diseases. There exists an urgent need to
define the structural and catalytic foundations of protein acylation by GOAT, Hhat, and PORCN in order to
understand the molecular basis of their diverse biological roles. The objective in this application is to define the
structural and chemical basis for transmembrane protein acylation by a membrane O-acyltransferase.
The studies proposed herein will develop a molecular-level structural model of GOAT verified and supported
by chemical and biochemical studies. Supported by strong preliminary studies, our objective will be pursued in
the following three Specific Aims: 1) Define the acyl donor and ghrelin binding sites within human GOAT
(hGOAT); 2) Determine the hGOAT catalytic mechanism, and 3) Identify inhibitor binding sites within hGOAT. In
the first Aim, an hGOAT structural model generated by bioinformatic analysis coupled with computational
modeling will guide studies to identify the acyl donor and ghrelin binding sites within hGOAT, with the ultimate
goal of defining how hGOAT accomplishes the topologically challenging transmembrane octanoylation of ghrelin.
In the second Aim, structure-guided mutagenesis and mechanistic probes will reveal the location and
composition of the active site responsible for ghrelin acylation. In the third Aim, inhibitor binding sites within
hGOAT will be identified using cysteine-reactive chemical probes and computational docking studies. This
proposal is innovative because it represents a new and substantive departure from the standard approaches for
investigating the structure and mechanism of membrane-bound enzymes. Our work will establish a novel
powerful and general approach for investigating structurally intractable membrane proteins. The proposed
research is significant because it will generate the first structure of an MBOAT enzyme family protein
acyltransferase while providing insight into the structures and catalytic strategies of MBOAT family members,
which will advance the exploitation of acylated secreted proteins as therapeutic targets for human diseases.
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SYNTHESIS OF AMPHIDINOLIDE P USING RUTHENIUM CATALYSIS
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批准号:6377928
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项目类别:
-
资助金额:$3.48万
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财政年份:2001
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负责人:John Daniel Chisholm
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依托单位:
SYNTHESIS OF AMPHIDINOLIDE P USING RUTHENIUM CATALYSIS
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批准号:6136304
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项目类别:
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资助金额:$3.09万
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财政年份:2000
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负责人:John Daniel Chisholm
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依托单位:
海外基金