Expression and purification of the full murine NPM2 and study of its interaction with protamines and histones

Katherine Ellard; Jason J Serpa; Evgeniy V Petrotchenko; Christoph H Borchers; Juan Ausió

doi:10.1016/j.bbrep.2016.04.002

Expression and purification of the full murine NPM2 and study of its interaction with protamines and histones

Biochem Biophys Rep. 2016 Apr 8:6:165-171. doi: 10.1016/j.bbrep.2016.04.002. eCollection 2016 Jul.

Authors

Katherine Ellard¹, Jason J Serpa^{1

2}, Evgeniy V Petrotchenko^{1

2}, Christoph H Borchers^{1

2}, Juan Ausió¹

Affiliations

¹ Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada V8W 3P6.
² Genome British Columbia Proteomics Centre, University of Victoria, Victoria, BC, Canada V8Z 7X8.

Abstract

Mouse nucleoplasmin M.NPM2 was recombinantly expressed and the protein consisting of the complete sequence was purified and characterized. Similar to its Xenopus laevis X.NPM2 counterpart, the protein forms stable pentameric complexes and exhibits an almost undistinguishable hydrodynamic ionic strength-dependent unfolding behavior. The interaction of N.PM2 with histones and mouse P1/P2 protamines revealed that these chromosomal proteins bind preferentially to the distal part of the nucleoplasmin pentamer. Moreover, the present work highlights the critical role played by histones H2B and H4 in the association of the histone H2A-H2B dimers and histone octamer with nucleoplasmin.