Chapter title |
Characterization of Protein Complexes Using Chemical Cross-Linking Coupled Electrospray Mass Spectrometry
|
---|---|
Chapter number | 85 |
Book title |
Tissue Proteomics
|
Published in |
Methods in molecular biology, October 2017
|
DOI | 10.1007/7651_2017_85 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7852-6, 978-1-4939-7854-0
|
Authors |
Timothy D. Cummins, Gopal P. Sapkota, Cummins, Timothy D., Sapkota, Gopal P. |
Abstract |
Identification and characterization of large protein complexes is a mainstay of biochemical toolboxes. Utilization of cross-linking chemicals can facilitate the capture and identification of transient or weak interactions of a transient nature (Huang and Kim, PloS One 8:e61430, 2013; Gao et al., J Vis Exp doi: 10.3791/51387, 2014). Here we describe a detailed methodology for a cell culture-based proteomic approach. We describe the generation of cells stably expressing green fluorescent protein (GFP)-tagged proteins under the tetracycline-inducible promoter and subsequent proteomic analysis of GFP-interacting proteins. We include a list of proteins that were identified as interactors of GFP. |
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