2. Academic Validation
  3. Purification of Native Acetyl CoA Carboxylase From Mammalian Cells

Purification of Native Acetyl CoA Carboxylase From Mammalian Cells

  • Bio Protoc. 2025 Feb 20;15(4):e5221. doi: 10.21769/BioProtoc.5221.
Yaxue Sun 1 2 3,  Jiachen Li 1 2 3,  Lianmei Zhao 1 2,  Hongtao Zhu 3 4
Affiliations

Affiliations

  • 1 Hebei Provincial Key Laboratory of Tumour Prevention and Precision Diagnosis and Treatment, Shijiazhuang, Hebei, China.
  • 2 Medical School, He'bei University, Shijiazhuang, Hebei, China.
  • 3 Beijing National Laboratory for Condensed Matter Physics and Laboratory of Soft Matter Physics, Institute of Physics, Beijing, China.
  • 4 University of Chinese Academy of Sciences, Beijing, China.
PMID: 40028013 DOI: 10.21769/BioProtoc.5221
Abstract

Fatty acid (FA) biosynthesis is a crucial cellular process that converts nutrients into metabolic intermediates necessary for membrane biosynthesis, energy storage, and the production of signaling molecules. Acetyl-CoA Carboxylase (ACACA) plays a pivotal catalytic role in both fatty acid synthesis and oxidation. This cytosolic enzyme catalyzes the carboxylation of acetyl-CoA to malonyl-CoA, which represents the first and rate-limiting step in de novo fatty acid biosynthesis. In this study, we developed a rapid and effective purification scheme for separating human ACACA without any exogenous affinity tags, providing researchers with a novel method to obtain human ACACA in its native form. Key features • Detailed protocol for the purification of native ACACA. • ACACA is biotinylated in mammalian cells. Graphical overview.

Keywords

ACACA; Biotinylated; Endogenous; Fatty acids (FAs); Tumors.

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