Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/97416
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Type: Journal article
Title: Eukaryotic expression, purification and structure/function analysis of native, recombinant CRISP3 from human and mouse
Author: Volpert, M.
Mangum, J.
Jamsai, D.
D'Sylva, R.
O'Bryan, M.
McIntyre, P.
Citation: Scientific Reports, 2014; 4(1):4217-1-4217-7
Publisher: Nature Publishing Group
Issue Date: 2014
ISSN: 2045-2322
2045-2322
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Responsibility: 
Marianna Volpert, Jonathan E. Mangum, Duangporn Jamsai, Rebecca D’Sylva, Moira K. O’Bryan & Peter McIntyre
Abstract: While the Cysteine-Rich Secretory Proteins (CRISPs) have been broadly proposed as regulators of reproduction and immunity, physiological roles have yet to be established for individual members of this family. Past efforts to investigate their functions have been limited by the difficulty of purifying correctly folded CRISPs from bacterial expression systems, which yield low quantities of correctly folded protein containing the eight disulfide bonds that define the CRISP family. Here we report the expression and purification of native, glycosylated CRISP3 from human and mouse, expressed in HEK 293 cells and isolated using ion exchange and size exclusion chromatography. Functional authenticity was verified by substrate-affinity, native glycosylation characteristics and quaternary structure (monomer in solution). Validated protein was used in comparative structure/function studies to characterise sites and patterns of N-glycosylation in CRISP3, revealing interesting inter-species differences.
Keywords: Cell Line
Rights: This work is licensed under a Creative Commons Attribution- NonCommercial-ShareAlike 3.0 Unported license. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0
DOI: 10.1038/srep04217
Grant ID: http://purl.org/au-research/grants/nhmrc/606562
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