Membrane-SPINE: An improved method to identify protein-protein interaction partners of membrane proteins in vivo

• Published in: Proteomics (Weinheim) Vol. 11; no. 10; pp. 2124 - 2128
• Main Authors: Müller, Volker S., Jungblut, Peter R., Meyer, Thomas F., Hunke, Sabine
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.05.2011; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Subjects: Affinity Labels; Bacterial Proteins - metabolism; Cloning, Molecular; Formaldehyde; Immunoblotting; Mass Spectrometry; Membrane proteins; Membrane Proteins - metabolism; Membrane-Strep-tagged protein interaction experiment; MS-analysis; Protein Binding; Protein Interaction Mapping - methods; Protein-protein interaction; Proteins; Proteins - analysis; Proteins - chemistry; Proteins - metabolism; Proteomics - methods; Salmonella enterica serovar Typhimurium; Salmonella typhimurium; Streptavidin - chemistry; Streptavidin - metabolism; Technology
• ISSN: 1615-9853; 1615-9861; 1615-9861
• DOI: 10.1002/pmic.201000558

Membrane proteins are crucial for many essential cellular processes. As membrane proteins function in complexes, methods to detect and to characterize membrane protein–protein interactions are undoubtedly required. Therefore, we developed the “Membrane‐Strep‐tagged protein interaction experiment” (Membrane‐SPINE) that combines the specific purification of a Strep‐tagged membrane protein with the reversible fixation of protein complexes by formaldehyde cross‐linking. In combination with MS analysis, we suggest Membrane‐SPINE as a powerful tool to identify unknown interaction partners of membrane proteins in vivo.