Expression, purification, and characterization of c-FLIP tandem death effector domains from Escherichia coli

Zhi-Qiang Bai; Kaifeng Hu

doi:10.1016/j.pep.2022.106168

Expression, purification, and characterization of c-FLIP tandem death effector domains from Escherichia coli

Protein Expr Purif. 2022 Dec:200:106168. doi: 10.1016/j.pep.2022.106168. Epub 2022 Sep 6.

Authors

Zhi-Qiang Bai¹, Kaifeng Hu²

Affiliations

¹ Innovative Institute of Chinese Medicine and Pharmacy, University of Traditional Chinese Medicine, Chengdu, 611137, China; College of Science, Yunnan Agricultural University, Kunming, 650201, China; State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, 650201, China.
² Innovative Institute of Chinese Medicine and Pharmacy, University of Traditional Chinese Medicine, Chengdu, 611137, China; State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, 650201, China. Electronic address: kaifenghu@cdutcm.edu.cn.

PMID: 36084903
DOI: 10.1016/j.pep.2022.106168

Abstract

Cellular FLICE-like inhibitory protein (c-FLIP) regulates extrinsic apoptosis by controlling procaspase-8 activation through its tandem N-terminal death effector domains (DEDs). Here, we present the expression and purification of c-FLIP tandem DEDs (tDED) from Escherichia coli. We observed that the c-FLIP^tDED maintains monomeric form under near-physiological pH condition in vitro. Our results also reveal a significant correlation between the pH conditions and the structure of c-FLIP^tDED (F114A). The described methods and results would be helpful for follow-up study on the structural and functional of c-FLIP.

Keywords: Death effector domains; Escherichia coli; Expression; Purification; c-FLIP.

Publication types

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

MeSH terms

Apoptosis
CASP8 and FADD-Like Apoptosis Regulating Protein* / genetics
CASP8 and FADD-Like Apoptosis Regulating Protein* / metabolism
Caspase 8 / metabolism
Death Effector Domain*
Escherichia coli / genetics
Escherichia coli / metabolism
Follow-Up Studies

Substances

CASP8 and FADD-Like Apoptosis Regulating Protein
Caspase 8