Human Adenine Nucleotide Translocase (ANT) Modulators Identified by High-Throughput Screening of Transgenic Yeast

被引：13

作者：

Zhang, Yujian ^{[1
]}

Tian, Defeng ^{[2
]}

Matsuyama, Hironori ^{[1
]}

Hamazaki, Takashi ^{[3
]}

Shiratsuchi, Takayuki ^{[1
]}

Terada, Naohiro ^{[4
]}

Hook, Derek J. ^{[2
]}

Walters, Michael A. ^{[2
]}

Georg, Gunda I. ^{[2
]}

Hawkinson, Jon E. ^{[2
]}

机构：

[1] Otsuka Maryland Med Labs Inc, Rockville, MD USA

[2] Univ Minnesota, Inst Therapeut Discovery & Dev, 717 Delaware St, Minneapolis, MN 55414 USA

[3] Osaka City Univ, Grad Sch Med, Dept Pediat, Abeno Ku, Osaka 558, Japan

[4] Univ Florida, Coll Med, Dept Pathol, Gainesville, FL USA

来源：

JOURNAL OF BIOMOLECULAR SCREENING | 2016年 / 21卷 / 04期

基金：

美国国家卫生研究院;

关键词：

adenine nucleotide translocase; high throughput screening; yeast mitochondria; Lactococcus lactis; ADP/ATP exchange; MITOCHONDRIAL ADP/ATP CARRIER; TRANSPORT; CHELERYTHRINE; INHIBITION;

D O I：

10.1177/1087057115624637

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Transport of ADP and ATP across mitochondria is one of the primary points of regulation to maintain cellular energy homeostasis. This process is mainly mediated by adenine nucleotide translocase (ANT) located on the mitochondrial inner membrane. There are four human ANT isoforms, each having a unique tissue-specific expression pattern and biological function, highlighting their potential as drug targets for diverse clinical indications, including male contraception and cancer. In this study, we present a novel yeast-based high-throughput screening (HTS) strategy to identify compounds inhibiting the function of ANT. Yeast strains generated by deletion of endogenous proteins with ANT activity followed by insertion of individual human ANT isoforms are sensitive to cell-permeable ANT inhibitors, which reduce proliferation. Screening hits identified in the yeast proliferation assay were characterized in ADP/ATP exchange assays employing recombinant ANT isoforms expressed in isolated yeast mitochondria and Lactococcus lactis as well as by oxygen consumption rate in mammalian cells. Using this approach, closantel and CD437 were identified as broad-spectrum ANT inhibitors, whereas leelamine was found to be a modulator of ANT function. This yeast knock-out/knock-in screening strategy is applicable to a broad range of essential molecular targets that are required for yeast survival.

引用

页码：381 / 390

页数：10

共 27 条

[1] Adenine nucleotide translocator mediates the mitochondrial membrane permeabilization induced by lonidamine, arsenite and CD437 [J].