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

• DOI: 10.1016/j.xpro.2022.101137
• Publication Date: 2022-01-28T00:30:44Z
• Authors: Lubos Cipak; Tomas Selicky; Jan Jurcik; Ingrid Cipakova; Michaela Osadska; Veronika Lukacova; Peter Barath; Juraj Gregan
• Keywords: 0301 basic medicine; Science (General); Tandem Affinity Purification; Proteins; Cell Biology; Mass Spectrometry; Q1-390; 03 medical and health sciences; Model Organisms; Protein Biochemistry; Schizosaccharomyces; Protein expression and purification; Protocol

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).