Work

Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro

Year: 2024

Type: article

Abstract: -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.

Source: BIO-PROTOCOL

DOI: 10.21769/bioprotoc.5095

Authors: Anup Parchure, Julia von Blume

Institution: Yale University

Language: English

Topic: Cellular transport and secretion

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: diamond

Funders: W. M. Keck Foundation, Yale University, National Institute of General Medical Sciences, National Institute of Diabetes and Digestive and Kidney Diseases

Awards: Coordinating Unit for the Innovative Science Accelerator Program, Mechanism of secretory cargo sorting at the trans-Golgi Network (TGN), Yale Diabetes Research Center, Vesicle Translocation and the Metabolic Syndrome