Administrative Supplement to Characterizing proteasome-substrate interactions by mass spectrometry proteomics
Administrative Supplement to Characterizing proteasome-substrate interactions by mass spectrometry proteomics
批准号:
10388694
负责人:
DAVID E. CLEMMER
金额:
$5.0万
依托单位国家:
美国
项目类别:
财政年份:
2020
资助国家:
美国
项目状态:
未结题
起止时间:
2020-07-01 至 2025-03-31
关键词:
Administrative SupplementBiochemicalBiologicalBiological AssayChargeConsensusDetectionDevicesDissociationElectron TransportIndividualIonsMass Spectrum AnalysisMethodologyMethodsModelingPatternPeptidesPolyubiquitinProteinsProteomicsPublishingReportingResearch Project GrantsScienceSignal TransductionSubstrate InteractionUbiquitinYeastscrosslinkdesignexperimental studyimprovedion sourcemulticatalytic endopeptidase complexnovelreceptorsuccesstransmission process
中文摘要
项目总结
英文摘要
PROJECT SUMMARY
We are proposing to purchase a Thermo FAIMS Pro Orbitrap Ion Source that will attach to the front
of a Lumos Orbitrap and be used in our ubiquitin-proteasome cross-linking experiments. Our research
project involves probing interactions between polyubiquitin chains, model proteasome substrates and
the yeast proteasome. Cross-linking mass spectrometry is being applied to discover novel ubiquitin
receptors and to better understand how tagged substrates can impact proteasome function. It is crucial
that cross-links are identified with high confidence because of the complexity of the biochemical assays
that build on these results. Since there is limited consensus in this field as to what constitutes an
unambiguous cross-link identification, we are exerting considerable effort to improve cross-linking
observation and interpretation methodology. By combining electron transfer dissociation, collision
induced dissociation, and high energy collision induced dissociation of precursor ions we have recently
reported how the credibility of cross-link identifications can be enhanced. Crucial to the success of this
approach is to have intense precursor ion signals. The Thermo FAIMS Pro Orbitrap Ion Source is
designed to increase the transmission of highly-charged ions such as those associated with cross-
linked peptides and to discriminate against background species such as individual peptides. In fact, it
has already demonstrated success in cross-linking mass spectrometry experiments. We will use this
device to discover and confirm cross-links involving polyubiquitin and the yeast proteasome and by
doing so will achieve both analytical and biological advances. Progress in our ability to recognize and
conclusively identify cross-linked peptides has transformative potential for elucidating some of the most
important and challenging issues in protein science. Specifically, our cross-linking proteomics
experiments will provide candidates for novel substrate receptors on the proteasome and will identify
the sizes and linkage patterns in polyubiquitin that are most effective for recognition by the proteasome.
In summary, this project should result in significant advances in cross-linking methodology and in our
understanding of how the proteasome recognizes and digests substrates.
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Characterizing proteasome-substrate interactions by mass spectrometry proteomics
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批准号:10200097
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项目类别:
-
资助金额:$34.18万
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财政年份:2020
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负责人:DAVID E. CLEMMER
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依托单位:
Characterizing proteasome-substrate interactions by mass spectrometry proteomics
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批准号:10377447
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项目类别:
-
资助金额:$34.15万
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财政年份:2020
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负责人:DAVID E. CLEMMER
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依托单位:
Developing High-Resolution Ion Mobility Spectrometry-Charge Detection-Mass Spectrometry for Rapid Analysis in the Megadalton to Gigadalton Regime
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批准号:10061629
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项目类别:
-
资助金额:$48.77万
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财政年份:2018
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负责人:DAVID E. CLEMMER
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依托单位:
Developing High-Resolution Ion Mobility Spectrometry-Charge Detection-Mass Spectrometry for Rapid Analysis in the Megadalton to Gigadalton Regime
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批准号:10295181
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项目类别:
-
资助金额:$50.29万
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财政年份:2018
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负责人:DAVID E. CLEMMER
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依托单位:
Development of high resolution mobility measurements for structural biology
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批准号:9383630
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项目类别:
-
资助金额:$47.77万
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财政年份:2017
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负责人:DAVID E. CLEMMER
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依托单位:
New proteome techniques: mapping adult D. Melanogaster
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批准号:9146961
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项目类别:
-
资助金额:$29.12万
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财政年份:2015
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负责人:DAVID E. CLEMMER
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依托单位:
New proteome techniques: mapping adult D. Melanogaster
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批准号:9009178
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项目类别:
-
资助金额:$25.41万
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财政年份:2015
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负责人:DAVID E. CLEMMER
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依托单位:
2011 Biological Molecules in the Gas Phase and in Solution GRC
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批准号:8193187
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项目类别:
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资助金额:$0.5万
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财政年份:2011
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负责人:DAVID E. CLEMMER
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依托单位:
Developing high-throughput IMS-MS and IMS-IMS-MS techniques for glycomics analysi
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批准号:7887486
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项目类别:
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资助金额:$30.24万
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财政年份:2010
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负责人:DAVID E. CLEMMER
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依托单位:
Developing high-throughput IMS-MS and IMS-IMS-MS techniques for glycomics analysi
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批准号:8306187
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项目类别:
-
资助金额:$28.93万
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财政年份:2010
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负责人:DAVID E. CLEMMER
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依托单位:
Developing high-throughput IMS-MS and IMS-IMS-MS techniques for glycomics analysi
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批准号:8473881
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项目类别:
-
资助金额:$27.89万
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财政年份:2010
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负责人:DAVID E. CLEMMER
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依托单位:
Developing high-throughput IMS-MS and IMS-IMS-MS techniques for glycomics analysi
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批准号:8078967
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项目类别:
-
资助金额:$27.49万
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财政年份:2010
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负责人:DAVID E. CLEMMER
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依托单位:
New Ion Mobility/Photodissociation Techniques for Analyzing Glycans
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批准号:7813541
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项目类别:
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资助金额:$49.63万
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财政年份:2009
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负责人:DAVID E. CLEMMER
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依托单位:
New Ion Mobility/Photodissociation Techniques for Analyzing Glycans
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批准号:7937881
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项目类别:
-
资助金额:$45.89万
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财政年份:2009
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负责人:DAVID E. CLEMMER
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依托单位:
CORE 3: MULTIDIMENSIONAL TECHNIQUES INVOLVING ION MOBILITY SEPARATIONS
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批准号:7724559
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项目类别:
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资助金额:$16.0万
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财政年份:2007
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负责人:DAVID E. CLEMMER
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依托单位:
CORE 3: MULTIDIMENSIONAL TECHNIQUES INVOLVING ION MOBILITY SEPARATIONS
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批准号:7602914
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项目类别:
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资助金额:$25.49万
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财政年份:2007
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负责人:DAVID E. CLEMMER
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依托单位:
CORE 3: MULTIDIMENSIONAL TECHNIQUES INVOLVING ION MOBILITY SEPARATIONS
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批准号:7359153
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项目类别:
-
资助金额:$17.57万
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财政年份:2006
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负责人:DAVID E. CLEMMER
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依托单位:
New proteome technologies: mapping adult D. melanogaster
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批准号:7415048
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项目类别:
-
资助金额:$24.48万
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财政年份:2005
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负责人:DAVID E. CLEMMER
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依托单位:
New proteome technologies: mapping adult D. melanogaster
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批准号:7224853
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项目类别:
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资助金额:$24.9万
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财政年份:2005
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负责人:DAVID E. CLEMMER
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依托单位:
New proteome technologies: mapping adult D. melanogaster
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批准号:6922561
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项目类别:
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资助金额:$23.03万
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财政年份:2005
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负责人:DAVID E. CLEMMER
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依托单位:
海外基金