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

B. Kuropka, N. Royla, C.Freund, E. Krause – 2014

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.

Titel
Sortase A mediated site-specific immobilization for identification of protein interactions in affinity purification–mass spectrometry experiments
Verfasser
B. Kuropka, N. Royla, C.Freund, E. Krause
Datum
2014
Kennung
10.1002/pmic.201400395
Zitierweise
Proteomics, 2014, 15, 1230–1234
Art
Text
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