Expression and Purification of Arabidopsis Transmembrane Protein BCM1 in Saccharomyces cerevisiae

Expression and Purification of Arabidopsis Transmembrane Protein BCM1 in Saccharomyces cerevisiae

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Heterologous expression and purification of transmembrane proteins have remained a challenge for decades hampering detailed biochemical and structural characterization of key enzymes and their interacting regulators in multiple metabolic pathways.An in-depth study on the newly identified Arabidopsis thaliana integral opi the color that keeps on giving membrane protein BALANCE OF CHLOROPHYLL METABOLISM 1 (BCM1) showed a stimulatory effect of the BCM1 on magnesium chelatase, the first enzyme of chlorophyll biosynthesis, through interaction with the GENOMES UNCOUPLED 4 (Wang et al., 2020).Here, we report a detailed and optimized method for heterologous expression and purification of borstlist självhäftande His-tagged BCM1 in Saccharomyces cerevisiae.Following this method, we obtained native BCM1 used for in vitro enzymatic assay of magnesium chelatase (Wang et al.

, 2020).Currently, the crystallization studies of the BCM1 are underway.This protocol could be adapted to purify BCM1-like transmembrane proteins from eukaryotic organisms for enzymatic and structural studies.