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Chromogranin B Purification for Condensate Formation and Client Partitioning
Assays In Vitro.
Authors
Parchure A, Von Blume J
Submitted By
Julia von Blume on 12/16/2024
Status
Published
Journal
Bio-protocol
Year
2024
Date Published
Volume : Pages
14 : e5095
PubMed Reference
39512880
Abstract
Chromogranin B and other members of the granin protein family form condensates
that recruit clients like proinsulin. The condensation in the lumen of
trans-Golgi network (TGN) is critical for the biogenesis of secretory granules.
Here, we describe a protocol to purify the tagged version of chromogranin B
close to its native form at the TGN, which can then be utilized for
microscopy-based assays to monitor condensate formation in vitro and client
partitioning depending on the material properties of chromogranin B assemblies.
Key features • First instance of purification of full-length and tagged version
of members of the chromogranin family of proteins. • Allows purification of
proteins with post-translational modifications that are acquired en route in the
secretory pathway, thus closely resembling their native form at the TGN.
Investigators with authorship
Name
Institution
Julia von Blume
Yale University
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