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Improved elution strategy and new monoclonal anti-biotin antibody for LC-MS/MS characterization of protein biotinylation sites.
Zhu, Yiying.
Afiliación
  • Zhu Y; Tsinghua University and Cell Signaling Technology Inc, Chemistry Department, Tsinghua University, Beijing, 100084, China.
Biochem Biophys Rep ; 38: 101711, 2024 Jul.
Article en En | MEDLINE | ID: mdl-38681669
ABSTRACT
Biotin labeling in combination with mass spectrometry has been widely applied in large-scale biological studies, such as determination of protein partners, protein subcellular localization, and protein post-translational modifications. Previous studies have shown that immunoaffinity enrichment is a better method than streptavidin/avidin purification for site-specific studies of biotinylated molecules. In this study, we made a crucial improvement to the elution phase of the immunoaffinity enrichment step for biotinylated peptides, which involves the addition of a highly organic solvent, and developed a monoclonal anti-biotin antibody that improved the identification number for biotinylated peptides. We then demonstrated its application in the characterization of protein interaction sites for the ß2 adrenergic receptor (ß2AR) by proximity labeling in living cells. Our research provides an improved and reproducible immunoaffinity enrichment method for site-specific biotin-related research.
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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Idioma: En Revista: Biochem Biophys Rep Año: 2024 Tipo del documento: Article País de afiliación: China

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Idioma: En Revista: Biochem Biophys Rep Año: 2024 Tipo del documento: Article País de afiliación: China