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Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro
Anup Parchure1,2, Julia Von Blume2
1Section of Endocrinology, Department of Internal Medicine, Yale School of Medicine, New Haven, CT, USA.
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.

