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Tandem affinity purification protocol for isolation of protein complexes from Schizosaccharomyces pombe

Many cellular processes require the activities of complex molecular machines composed of several protein subunits. Insights into these systems can be gained by isolation of protein complexes followed by in vitro analyses determining the identity, posttranslational modifications, and interactions amo...

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Detalles Bibliográficos
Autores principales: Cipak, Lubos, Selicky, Tomas, Jurcik, Jan, Cipakova, Ingrid, Osadska, Michaela, Lukacova, Veronika, Barath, Peter, Gregan, Juraj
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8808283/
https://www.ncbi.nlm.nih.gov/pubmed/35128479
http://dx.doi.org/10.1016/j.xpro.2022.101137
Descripción
Sumario:Many cellular processes require the activities of complex molecular machines composed of several protein subunits. Insights into these systems can be gained by isolation of protein complexes followed by in vitro analyses determining the identity, posttranslational modifications, and interactions among proteins. Here, we present a protocol for tandem affinity purification (TAP) of protein complexes from the fission yeast Schizosaccharomyces pombe. The protocol employs cells expressing C-terminally TAP-tagged proteins and is suitable for the analysis of purified proteins by mass spectrometry. For complete information on the use and execution of this protocol, please refer to Cipakova et al. (2019).