DC Field | Value | Language |
---|---|---|
dc.contributor.author | Sohn, Chang Ho | ko |
dc.contributor.author | Agnew, Heather D. | ko |
dc.contributor.author | Lee, J. Eugene | ko |
dc.contributor.author | Sweredoski, Michael J. | ko |
dc.contributor.author | Graham, Robert L. J. | ko |
dc.contributor.author | Smith, Geoffrey T. | ko |
dc.contributor.author | Hess, Sonja | ko |
dc.contributor.author | Czerwieniec, Gregg | ko |
dc.contributor.author | Loo, Joseph A. | ko |
dc.contributor.author | Heath, James R. | ko |
dc.contributor.author | Deshaies, Raymond J. | ko |
dc.contributor.author | Beauchamp, J. L. | ko |
dc.date.accessioned | 2024-06-03T08:01:10Z | - |
dc.date.available | 2024-06-03T08:01:10Z | - |
dc.date.created | 2024-06-03 | - |
dc.date.created | 2024-06-03 | - |
dc.date.created | 2024-06-03 | - |
dc.date.issued | 2012-03 | - |
dc.identifier.citation | ANALYTICAL CHEMISTRY, v.84, no.6, pp.2662 - 2669 | - |
dc.identifier.issn | 0003-2700 | - |
dc.identifier.uri | http://hdl.handle.net/10203/319609 | - |
dc.description.abstract | We present novel homobifunctional amine-reactive clickable cross-linkers (CXLs) for investigation of three-dimensional protein structures and protein-protein interactions (PPIs). CXLs afford consolidated advantages not previously available in a simple cross-linker, including (1) their small size and cationic nature at physiological pH, resulting in good water solubility and cell-permeability, (2) an allcyne group for bio-orthogonal conjugation to affinity tags via the click reaction for enrichment of cross-linked peptides, (3) a nucleophilic displacement reaction involving the 1,2,3-triazole ring formed in the click reaction, yielding a lock-mass reporter ion for only clicked peptides, and (4) higher charge states of cross-linked peptides in the gas-phase for augmented electron transfer dissociation (ETD) yields. Ubiquitin, a lysine-abundant protein, is used as a model system to demonstrate structural studies using CXLs. To validate the sensitivity of our approach, biotin-azide labeling and subsequent enrichment of cross-linked peptides are performed for cross-linked ubiquitin digests mixed with yeast cell lysates. Cross-linked peptides are detected and identified by collision induced dissociation (CID) and ETD with linear quadrupole ion trap (LTQ)-Fourier transform ion cyclotron resonance (FTICR) and LTQ:Orbitrap mass spectrometers. The application of CXLs to more complex systems (e.g., in vivo cross-linking) is illustrated by Western blot detection of Cull complexes including known binders, Cand1 and Skp2, in HEK 293 cells, confirming good water solubility and cell-permeability. | - |
dc.language | English | - |
dc.publisher | AMER CHEMICAL SOC | - |
dc.title | Designer Reagents for Mass Spectrometry-Based Proteomics: Clickable Cross-Linkers for Elucidation of Protein Structures and Interactions | - |
dc.type | Article | - |
dc.identifier.wosid | 000301634500008 | - |
dc.identifier.scopusid | 2-s2.0-84863388262 | - |
dc.type.rims | ART | - |
dc.citation.volume | 84 | - |
dc.citation.issue | 6 | - |
dc.citation.beginningpage | 2662 | - |
dc.citation.endingpage | 2669 | - |
dc.citation.publicationname | ANALYTICAL CHEMISTRY | - |
dc.identifier.doi | 10.1021/ac202637n | - |
dc.contributor.localauthor | Sohn, Chang Ho | - |
dc.contributor.nonIdAuthor | Agnew, Heather D. | - |
dc.contributor.nonIdAuthor | Lee, J. Eugene | - |
dc.contributor.nonIdAuthor | Sweredoski, Michael J. | - |
dc.contributor.nonIdAuthor | Graham, Robert L. J. | - |
dc.contributor.nonIdAuthor | Smith, Geoffrey T. | - |
dc.contributor.nonIdAuthor | Hess, Sonja | - |
dc.contributor.nonIdAuthor | Czerwieniec, Gregg | - |
dc.contributor.nonIdAuthor | Loo, Joseph A. | - |
dc.contributor.nonIdAuthor | Heath, James R. | - |
dc.contributor.nonIdAuthor | Deshaies, Raymond J. | - |
dc.contributor.nonIdAuthor | Beauchamp, J. L. | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordPlus | TOP-DOWN APPROACH | - |
dc.subject.keywordPlus | SELECTIVE ENRICHMENT | - |
dc.subject.keywordPlus | LIVING CELLS | - |
dc.subject.keywordPlus | LINKING | - |
dc.subject.keywordPlus | IDENTIFICATION | - |
dc.subject.keywordPlus | PEPTIDES | - |
dc.subject.keywordPlus | COMPLEXES | - |
dc.subject.keywordPlus | STRATEGY | - |
dc.subject.keywordPlus | PURIFICATION | - |
dc.subject.keywordPlus | DISSOCIATION | - |
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