Identification of Protein Complexes in Escherichia coli using Sequential Peptide Affinity Purification in Combination with Tandem Mass Spectrometry
Title:
Identification of Protein Complexes in Escherichia coli using Sequential Peptide Affinity Purification in Combination with Tandem Mass Spectrometry
Author:
Babu, Mohan
Personal Author:
Publication Information:
Cambridge, MA MyJoVE Corp 2016
Physical Description:
online resource (898 seconds)
Series:
Biology
General Note:
Title from resource description page
Abstract:
Affinity purification of tagged proteins in combination with mass spectrometry (APMS) is a powerful method for the systematic mapping of protein interaction networks and for investigating the mechanistic basis of biological processes. Here, we describe an optimized sequential peptide affinity (SPA) APMS procedure developed for the bacterium Escherichia coli that can be used to isolate and characterize stable multi-protein complexes to near homogeneity even starting from low copy numbers per cell.
Reading Level:
For undergraduate, graduate, and professional students
Subject Term:
Electronic Access:
https://www.jove.com/t/4057