La Trobe

Rapid purification method for human SFPQ by implementing zinc-induced polymerization

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posted on 2025-02-06, 02:19 authored by Yee Wa Lim, Mihwa LeeMihwa Lee
Splicing factor proline- and glutamine-rich (SFPQ) is an RNA-binding protein, playing significant roles in gene regulation and subnuclear body formation. Our recent serendipitous discovery showed that SFPQ binds zinc directly and forms an infinite polymer that is induced by zinc binding to the protein. The zinc-induced reversible polymerization has led us to exploit this property to develop a rapid purification strategy for SFPQ without the use of affinity tags. In combination with the variation of ionic strength for salting-out of SFPQ, the reversible zinc-induced precipitation of SFPQ reduced the purification time required to obtain pure SFPQ to a single day. The purified protein was subjected to the previously reported crystallization condition. The resulting crystals diffracted to 2.22 Å resolution, confirming the quality of SFPQ purified with this new rapid purification strategy.

Funding

This study was supported by the Australian Research Council Discovery Early Career Research Award (ARC DECRA) Fellowship (DE150101243) and Tracey Banivanua Mar Fellowship (La Trobe University, Melbourne, Australia) to ML.

History

Publication Date

2020-07-01

Journal

Protein Expression and Purification

Volume

171

Article Number

105626

Pagination

4p.

Publisher

Elsevier

ISSN

1046-5928

Rights Statement

© 2020 Elsevier Inc. All rights reserved. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/

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