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

Protein Expr Purif

Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria, 3086, Australia. Electronic address:

Published: July 2020

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.

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.pep.2020.105626DOI Listing

Publication Analysis

Top Keywords

rapid purification
12
sfpq
8
purification strategy
8
purification method
4
method human
4
human sfpq
4
sfpq implementing
4
implementing zinc-induced
4
zinc-induced polymerization
4
polymerization splicing
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!