Functional Characterization of the γ-Aminobutyric Acid Transporter from Mycobacterium smegmatis MC2 155 Reveals Sodium-Driven GABA Transport

被引：3

作者：

Pavic, Ana ^{[1
,2
]}

Ji, Yurui ^{[1
,3
,8
]}

Serafini, Agnese ^{[4
]}

Garza-Garcia, Acely ^{[4
]}

McPhillie, Martin J. ^{[5
]}

Holmes, Alexandra O. M. ^{[1
]}

de Carvalho, Luiz Pedro Sorio ^{[4
]}

Wang, Yingying ^{[3
]}

Bartlam, Mark ^{[6
]}

Goldman, Adrian ^{[1
,6
,7
]}

Postis, Vincent L. G. ^{[1
,2
]}

机构：

[1] Univ Leeds, Astbury Ctr Struct Mol Biol, Sch Biomed Sci, Leeds, W Yorkshire, England

[2] Leeds Beckett Univ, Fac Hlth & Social Sci, Biomed Res Grp, Leeds, W Yorkshire, England

[3] Nankai Univ, Coll Environm Sci & Engn, Tianjin, Peoples R China

[4] Francis Crick Inst, Mycobacterial Metab & Antibiot Res Lab, London, England

[5] Univ Leeds, Fac Engn & Phys Sci, Sch Chem, Leeds, W Yorkshire, England

[6] Nankai Univ, Coll Life Sci, Tianjin, Peoples R China

[7] Univ Helsinki, Mol & Integrat Biosci, Helsinki, Finland

[8] Tianjin Sino German Univ Appl Sci, New Energy Dept, Tianjin, Peoples R China

来源：

JOURNAL OF BACTERIOLOGY | 2021年 / 203卷 / 04期

基金：

美国国家科学基金会; 英国医学研究理事会; 英国生物技术与生命科学研究理事会; 英国惠康基金;

关键词：

GABA; membrane biology; mycobacteria; transporter; MULTIPLE SEQUENCE ALIGNMENT; MEMBRANE-PROTEINS; APC SUPERFAMILY; EXPRESSION; NITROGEN; GENE; PERMEASE; IDENTIFICATION; TUBERCULOSIS; METABOLISM;

D O I：

10.1128/JB.00642-20

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Characterizing the mycobacterial transporters involved in the uptake and/or catabolism of host-derived nutrients required by mycobacteria may identify novel drug targets against tuberculosis. Here, we identify and characterize a member of the amino acid-polyamine-organocation superfamily, a potential gamma-aminobutyric acid (GABA) transport protein, GabP, from Mycobacterium smegmatis. The protein was expressed to a level allowing its purification to homogeneity, and size exclusion chromatography coupled with multiangle laser light scattering (SEC-MALLS) analysis of the purified protein showed that it was dimeric. We showed that GabP transported gamma-aminobutyric acid both in vitro and when overexpressed in E. coli. Additionally, transport was greatly reduced in the presence of beta-alanine, suggesting it could be either a substrate or inhibitor of GabP. Using GabP reconstituted into proteoliposomes, we demonstrated that gamma-aminobutyric acid uptake is driven by the sodium gradient and is stimulated by membrane potential. Molecular docking showed that gamma-aminobutyric acid binds MsGabP, another Mycobacterium smegmatis putative GabP, and the Mycobacterium tuberculosis homologue in the same manner. This study represents the first expression, purification, and characterization of an active gamma-aminobutyric acid transport protein from mycobacteria. IMPORTANCE The spread of multidrug-resistant tuberculosis increases its global health impact in humans. As there is transmission both to and from animals, the spread of the disease also increases its effects in a broad range of animal species. Identifying new mycobacterial transporters will enhance our understanding of mycobacterial physiology and, furthermore, provides new drug targets. Our target protein is the gene product of msmeg_6196, annotated as GABA permease, from Mycobacterium smegmatis strain MC2 155. Our current study demonstrates it is a sodium-dependent GABA transporter that may also transport beta-alanine. As GABA may well be an essential nutrient for mycobacterial metabolism inside the host, this could be an attractive target for the development of new drugs against tuberculosis.

引用

页数：14