Nonprotein based enrichment method to analyze peptide cross-linking in protein complexes.
Anal Chem
; 81(17): 7149-59, 2009 Sep 01.
Article
em En
| MEDLINE
| ID: mdl-19642656
Cross-linking analysis of protein complexes and structures by tandem mass spectrometry (MS/MS) has advantages in speed, sensitivity, specificity, and the capability of handling complicated protein assemblies. However, detection and accurate assignment of the cross-linked peptides are often challenging due to their low abundance and complicated fragmentation behavior in collision-induced dissociation (CID). To simplify the MS analysis and improve the signal-to-noise ratio of the cross-linked peptides, we developed a novel peptide enrichment strategy that utilizes a cross-linker with a cryptic thiol group and using beads modified with a photocleavable cross-linker. The functional cross-linkers were designed to react with the primary amino groups in proteins. Human serum albumin was used as a model protein to detect intra- and intermolecular cross-linkages. Use of this protein-free selective retrieval method eliminates the contamination that can result from avidin-biotin based retrieval systems and simplifies data analysis. These features may make the method suitable to investigate protein-protein interactions in biological samples.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Peptídeos
/
Compostos de Sulfidrila
/
Albumina Sérica
/
Proteínas
/
Reagentes de Ligações Cruzadas
/
Espectrometria de Massas em Tandem
Tipo de estudo:
Evaluation_studies
Limite:
Humans
Idioma:
En
Revista:
Anal Chem
Ano de publicação:
2009
Tipo de documento:
Article
País de afiliação:
Estados Unidos