2.Discovery of the first macrolide antibiotic binding protein in Mycobacterium tuberculosis: a new antibiotic resistance drug target.
Qingqing ZHANG ; Huijuan LIU ; Xiang LIU ; Dunquan JIANG ; Bingjie ZHANG ; Hongliang TIAN ; Cheng YANG ; Luke W GUDDAT ; Haitao YANG ; Kaixia MI ; Zihe RAO
Protein & Cell 2018;9(11):971-975
3.Cryo-EM structures for the Mycobacterium tuberculosis iron-loaded siderophore transporter IrtAB.
Shan SUN ; Yan GAO ; Xiaolin YANG ; Xiuna YANG ; Tianyu HU ; Jingxi LIANG ; Zhiqi XIONG ; Yuting RAN ; Pengxuan REN ; Fang BAI ; Luke W GUDDAT ; Haitao YANG ; Zihe RAO ; Bing ZHANG
Protein & Cell 2023;14(6):448-458
The adenosine 5'-triphosphate (ATP)-binding cassette (ABC) transporter, IrtAB, plays a vital role in the replication and viability of Mycobacterium tuberculosis (Mtb), where its function is to import iron-loaded siderophores. Unusually, it adopts the canonical type IV exporter fold. Herein, we report the structure of unliganded Mtb IrtAB and its structure in complex with ATP, ADP, or ATP analogue (AMP-PNP) at resolutions ranging from 2.8 to 3.5 Å. The structure of IrtAB bound ATP-Mg2+ shows a "head-to-tail" dimer of nucleotide-binding domains (NBDs), a closed amphipathic cavity within the transmembrane domains (TMDs), and a metal ion liganded to three histidine residues of IrtA in the cavity. Cryo-electron microscopy (Cryo-EM) structures and ATP hydrolysis assays show that the NBD of IrtA has a higher affinity for nucleotides and increased ATPase activity compared with IrtB. Moreover, the metal ion located in the TM region of IrtA is critical for the stabilization of the conformation of IrtAB during the transport cycle. This study provides a structural basis to explain the ATP-driven conformational changes that occur in IrtAB.
Siderophores/metabolism*
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Iron/metabolism*
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Mycobacterium tuberculosis/metabolism*
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Cryoelectron Microscopy
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Adenosine Triphosphate/metabolism*
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ATP-Binding Cassette Transporters
4.Cryo-EM snapshots of mycobacterial arabinosyltransferase complex EmbB-AcpM.
Lu ZHANG ; Yao ZHAO ; Ruogu GAO ; Jun LI ; Xiuna YANG ; Yan GAO ; Wei ZHAO ; Sudagar S GURCHA ; Natacha VEERAPEN ; Sarah M BATT ; Kajelle Kaur BESRA ; Wenqing XU ; Lijun BI ; Xian'en ZHANG ; Luke W GUDDAT ; Haitao YANG ; Quan WANG ; Gurdyal S BESRA ; Zihe RAO
Protein & Cell 2020;11(7):505-517
Inhibition of Mycobacterium tuberculosis (Mtb) cell wall assembly is an established strategy for anti-TB chemotherapy. Arabinosyltransferase EmbB, which catalyzes the transfer of arabinose from the donor decaprenyl-phosphate-arabinose (DPA) to its arabinosyl acceptor is an essential enzyme for Mtb cell wall synthesis. Analysis of drug resistance mutations suggests that EmbB is the main target of the front-line anti-TB drug, ethambutol. Herein, we report the cryo-EM structures of Mycobacterium smegmatis EmbB in its "resting state" and DPA-bound "active state". EmbB is a fifteen-transmembrane-spanning protein, assembled as a dimer. Each protomer has an associated acyl-carrier-protein (AcpM) on their cytoplasmic surface. Conformational changes upon DPA binding indicate an asymmetric movement within the EmbB dimer during catalysis. Functional studies have identified critical residues in substrate recognition and catalysis, and demonstrated that ethambutol inhibits transferase activity of EmbB by competing with DPA. The structures represent the first step directed towards a rational approach for anti-TB drug discovery.
5.Cryo-EM structures of a prokaryotic heme transporter CydDC.
Chen ZHU ; Yanfeng SHI ; Jing YU ; Wenhao ZHAO ; Lingqiao LI ; Jingxi LIANG ; Xiaolin YANG ; Bing ZHANG ; Yao ZHAO ; Yan GAO ; Xiaobo CHEN ; Xiuna YANG ; Lu ZHANG ; Luke W GUDDAT ; Lei LIU ; Haitao YANG ; Zihe RAO ; Jun LI
Protein & Cell 2023;14(12):919-923
6.High-throughput screening identifies established drugs as SARS-CoV-2 PLpro inhibitors.
Yao ZHAO ; Xiaoyu DU ; Yinkai DUAN ; Xiaoyan PAN ; Yifang SUN ; Tian YOU ; Lin HAN ; Zhenming JIN ; Weijuan SHANG ; Jing YU ; Hangtian GUO ; Qianying LIU ; Yan WU ; Chao PENG ; Jun WANG ; Chenghao ZHU ; Xiuna YANG ; Kailin YANG ; Ying LEI ; Luke W GUDDAT ; Wenqing XU ; Gengfu XIAO ; Lei SUN ; Leike ZHANG ; Zihe RAO ; Haitao YANG
Protein & Cell 2021;12(11):877-888
A new coronavirus (SARS-CoV-2) has been identified as the etiologic agent for the COVID-19 outbreak. Currently, effective treatment options remain very limited for this disease; therefore, there is an urgent need to identify new anti-COVID-19 agents. In this study, we screened over 6,000 compounds that included approved drugs, drug candidates in clinical trials, and pharmacologically active compounds to identify leads that target the SARS-CoV-2 papain-like protease (PLpro). Together with main protease (M
Antiviral Agents/therapeutic use*
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Binding Sites
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COVID-19/virology*
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Coronavirus Papain-Like Proteases/metabolism*
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Crystallography, X-Ray
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Drug Evaluation, Preclinical
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Drug Repositioning
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High-Throughput Screening Assays/methods*
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Humans
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Imidazoles/therapeutic use*
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Inhibitory Concentration 50
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Molecular Dynamics Simulation
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Mutagenesis, Site-Directed
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Naphthoquinones/therapeutic use*
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Protease Inhibitors/therapeutic use*
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Protein Structure, Tertiary
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Recombinant Proteins/isolation & purification*
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SARS-CoV-2/isolation & purification*