Sortase A mediated site-specific immobilization for identification of protein interactions in affinity purification-mass spectrometry experiments

作者：Kuropka Benno; Royla Nadine; Freund Christian; Krause Eberhard^*

来源：Proteomics, 2015, 15(7): 1230-1234.

DOI：10.1002/pmic.201400395

• Mass spectrometry
• Protein interactions
• SILAC
• Sortase A
• Technology

Proteomics approaches using MS in combination with affinity purification have emerged as powerful tools to study protein-protein interactions. Here we make use of the specificity of sortase A transpeptidation reaction to prepare affinity matrices in which a protein bait is covalently linked to the matrix via a short C-terminal linker region. As a result of this site-directed immobilization, the bait remains functionally accessible to protein interactions. To apply this approach, we performed SILAC-based pull-down experiments and demonstrate the suitability of the approach.

• 出版日期2015-4