Abstract
Proteins act as executors for almost all kinds of cellular processes. The majority of proteins achieve their proper functions through interacting with other proteins. Knowing the binding partners of a protein is instrumental for understanding its function. The antibody pull-down method is a powerful and common approach to detect protein-protein interactions. Here, an antibody pull-down protocol is described for detecting protein-protein interactions in fission yeast Schizosaccharomyces pombe.
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
He H, Zhang S, Wang D, Hochwagen A, Li F (2016) Condensin promotes position effects within tandem DNA repeats via the RITS complex. Cell Rep 14(5):1018–1024. https://doi.org/10.1016/j.celrep.2016.01.006
Li F, Martienssen R, Cande WZ (2011) Coordination of DNA replication and histone modification by the Rik1-Dos2 complex. Nature 475(7355):244–248. https://doi.org/10.1038/nature10161
Li F, Huarte M, Zaratiegui M, Vaughn MW, Shi Y, Martienssen R, Cande WZ (2008) Lid2 is required for coordinating H3K4 and H3K9 methylation of heterochromatin and euchromatin. Cell 135(2):272–283. https://doi.org/10.1016/j.cell.2008.08.036
Gonzalez M, He H, Sun S, Li C, Li F (2013) Cell cycle-dependent deposition of CENP-A requires the Dos1/2-Cdc20 complex. Proc Natl Acad Sci U S A 110(2):606–611. https://doi.org/10.1073/pnas.1214874110
Dong Q, Yin FX, Gao F, Shen Y, Zhang F, Li Y, He H, Gonzalez M, Yang J, Zhang S, Su M, Chen YH, Li F (2016) Ccp1 homodimer mediates chromatin integrity by antagonizing CENP-A loading. Mol Cell 64(1):79–91. https://doi.org/10.1016/j.molcel.2016.08.022
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685
Kawashima SA, Tsukahara T, Langegger M, Hauf S, Kitajima TS, Watanabe Y (2007) Shugoshin enables tension-generating attachment of kinetochores by loading Aurora to centromeres. Genes Dev 21(4):420–435. https://doi.org/10.1101/gad.1497307
Lee CY, Stewart EV, Hughes BT, Espenshade PJ (2009) Oxygen-dependent binding of Nro1 to the prolyl hydroxylase Ofd1 regulates SREBP degradation in yeast. EMBO J 28(2):135–143. https://doi.org/10.1038/emboj.2008.271
Nikko E, Sullivan JA, Pelham HR (2008) Arrestin-like proteins mediate ubiquitination and endocytosis of the yeast metal transporter Smf1. EMBO Rep 9(12):1216–1221. https://doi.org/10.1038/embor.2008.199
Acknowledgments
We would like to thank past and present members of Li lab who contributed to the methods presented here, and thank Dr. Jinpu Yang for reading the manuscript. F.L. is a Pew Scholar in the Biomedical Sciences, supported by The Pew Charitable Trusts. This work was supported by National Institutes of Health grant R01GM106037 (to F.L.)., National Science Foundation grant MCB-1330557 (to F.L.), and the National Natural Science Foundation of China (NSFC) Grant 31728010 (to F.L.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Dong, Q., Li, F. (2018). Antibody Pull-Down Experiments in Fission Yeast. In: Singleton, T. (eds) Schizosaccharomyces pombe. Methods in Molecular Biology, vol 1721. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7546-4_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7546-4_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7545-7
Online ISBN: 978-1-4939-7546-4
eBook Packages: Springer Protocols