Protocol to quantify palmitoylation of cysteines in budding yeast

Yuqing Lei; Jiangli Zhu; Huihui Li; Eryan Kong; Kefeng Lu

doi:10.1016/j.xpro.2021.101035

Protocol to quantify palmitoylation of cysteines in budding yeast

STAR Protoc. 2021 Dec 15;3(1):101035. doi: 10.1016/j.xpro.2021.101035. eCollection 2022 Mar 18.

Authors

Yuqing Lei¹, Jiangli Zhu², Huihui Li¹, Eryan Kong³, Kefeng Lu¹

Affiliations

¹ Department of Pathology, West China Second University Hospital, State Key Laboratory of Biotherapy, Sichuan University, Chengdu 610041, China.
² Department of Urology, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and National Collaborative Innovation Center, Chengdu 610041, China.
³ Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang 453003, China.

Abstract

Palmitoylation is a special kind of lipid modification that targets proteins to membranes. This protocol introduces the acyl-biotin exchange (ABE) assay to determine the palmitoylation of protein cysteines in yeast. Palmitoylation is exchanged by biotinylated compounds so that the palmitoyl proteins can be affinity-purified for downstream assay by western blot. This protocol is easy to perform and can be applied to other biological sources with slight modifications. This protocol is limited to the detection of cysteine-based palmitoylation. For complete details on the use and execution of this profile, please refer to Lei et al. (2021).

Keywords: Cell Biology; Model Organisms; Protein Biochemistry; Protein expression and purification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blotting, Western
Cysteine / metabolism
Lipoylation*
Proteins / metabolism
Saccharomycetales* / metabolism

Substances

Proteins
Cysteine