Abstract
A method is described for the elucidation of protein-protein interactions using novel cross-linking reagents and mass spectrometry. The method incorporates (1) a modular solid-phase synthetic strategy for generating the cross-linking reagents, (2) enrichment and digestion of cross-linked proteins using microconcentrators, (3) mass spectrometric analysis of cross-linked peptides, and (4) comprehensive computational analysis of the cross-linking data. This integrated approach has been applied to the study of cross-linking between the components of the heterodimeric protein complex negative cofactor 2.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Acrylic Resins / chemistry
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Amino Acid Sequence
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Biotin / chemistry
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Cross-Linking Reagents / chemical synthesis
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Cross-Linking Reagents / chemistry*
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Glycine / chemistry
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Models, Molecular
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Molecular Sequence Data
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Peptide Fragments / chemistry
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Phosphoproteins / chemistry*
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Polyethylene Glycols / chemistry
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
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Sulfhydryl Compounds / chemistry
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Transcription Factors / chemistry*
Substances
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Acrylic Resins
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Cross-Linking Reagents
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Peptide Fragments
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Phosphoproteins
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Sulfhydryl Compounds
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Transcription Factors
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down-regulator of transcription 1
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Polyethylene Glycols
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Biotin
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polyacrylamide
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Glycine