Abstract
Co-immunoprecipitation is a classical method for the study of protein–protein and protein–DNA interactions, which is based on the specific recognition of antigens by antibodies (Sahr T, Buchrieser C: Methods Mol Biol 954:583–593, 2013). It is an effective method for determining the physiological interactions of two proteins under physiological conditions in intact cells. The protein–protein interactions identified by in vitro detection assays require further studies to confirm that they occur inside living cells. Fusing a small peptide tag, for instance, the FLAG-tag, to a protein of interest facilitates its co-immunoprecipitation using a commercially available anti-FLAG affinity gel. Compared with other methods for studying protein–protein interactions (such as pulldown assays and yeast two-hybrid screening), the protein–protein interactions identified by immunoprecipitation are generally an accurate reflection of the in vivo situation.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sahr T, Buchrieser C (2013) Co-immunoprecipitation: protein-RNA and protein-DNA interaction. Methods Mol Biol 954:583–593
Hoiczyk E, Blobel G (2001) Polymerization of a single protein of the pathogen Yersinia enterocolitica into needles punctures eukaryotic cells. Proc Natl Acad Sci U S A 98(8):4669–4674
Monlezun L, Liebl D, Fenel D, Grandjean T, Berry A, Schoehn G, Dessein R, Faudry E, Attree I (2015) PscI is a type III secretion needle anchoring protein with in vitro polymerization capacities. Mol Microbiol 96(2):419–436
Yang H, Tan Y, Zhang T, Tang L, Wang J, Ke Y, Guo Z, Yang X, Yang R, Du Z (2013) Identification of novel protein-protein interactions of Yersinia pestis type III secretion system by yeast two hybrid system. PLoS One 8(1):e54121
Cao SY, Liu WB, Tan YF, Yang HY, Zhang TT, Wang T, Wang XY, Song YJ, Yang RF, Du ZM (2017) An interaction between the inner rod protein YscI and the needle protein YscF is required to assemble the needle structure of the Yersinia type three secretion system. J Biol Chem 292(13):5488–5498
Du Z, Tan Y, Yang H, Qiu J, Qin L, Wang T, Liu H, Bi Y, Song Y, Guo Z et al (2009) Gene expression profiling of Yersinia pestis with deletion of lcrG, a known negative regulator for Yop secretion of type III secretion system. Int J Med Microbiol 299(5):355–366
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Nature Singapore Pte Ltd.
About this protocol
Cite this protocol
Cao, S., Du, Z. (2018). Co-immunoprecipitation Analysis for the Detection of Protein–Protein Interactions in Yersinia pestis . In: Yang, R. (eds) Yersinia Pestis Protocols. Springer Protocols Handbooks. Springer, Singapore. https://doi.org/10.1007/978-981-10-7947-4_15
Download citation
DOI: https://doi.org/10.1007/978-981-10-7947-4_15
Published:
Publisher Name: Springer, Singapore
Print ISBN: 978-981-10-7946-7
Online ISBN: 978-981-10-7947-4
eBook Packages: Springer Protocols