The Experts below are selected from a list of 156 Experts worldwide ranked by ideXlab platform
Pei Zhou - One of the best experts on this subject based on the ideXlab platform.
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Structure of the essential Haemophilus influenzae UDP-diacylglucosamine pyrophosphohydrolase LpXH in Lipid A biosynthesis
Nature Microbiology, 2016Co-Authors: Jae Cho, Chul-jin Lee, Jinshi Zhao, Hayley E. Young, Pei ZhouAbstract:In most Gram-negative pathogens, the hydrolysis of UDP-2,3-diacylglucosamine to generate Lipid X in Lipid A biosynthesis is catalysed by the membrane-associated enzyme LpXH. We report the crystal structure of LpXH in compleX with its product, Lipid X, unveiling a unique insertion lid above the conserved architecture of calcineurin-like phosphoesterases. This structure reveals elaborate interactions surrounding Lipid X and provides molecular insights into the substrate selectivity, catalysis and inhibition of LpXH. The structure of the LipidA diacylglucosamine pyrophosphohydrolase LpXH bound to its Lipid X substrate is presented.
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Structure of the essential Haemophilus influenzae UDP-diacylglucosamine pyrophosphohydrolase LpXH in Lipid A biosynthesis.
Nature microbiology, 2016Co-Authors: Jae Cho, Chul-jin Lee, Jinshi Zhao, Hayley E. Young, Pei ZhouAbstract:In most Gram-negative pathogens, the hydrolysis of UDP-2,3-diacylglucosamine to generate Lipid X in Lipid A biosynthesis is catalysed by the membrane-associated enzyme LpXH. We report the crystal structure of LpXH in compleX with its product, Lipid X, unveiling a unique insertion lid above the conserved architecture of calcineurin-like phosphoesterases. This structure reveals elaborate interactions surrounding Lipid X and provides molecular insights into the substrate selectivity, catalysis and inhibition of LpXH.
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Discovery of the Elusive UDP-Diacylglucosamine Hydrolase in the Lipid A Biosynthetic Pathway in Chlamydia trachomatis
mBio, 2016Co-Authors: Hayley E. Young, Jinshi Zhao, Jeffrey R. Barker, Ziqiang Guan, Raphael H. Valdivia, Pei ZhouAbstract:Constitutive biosynthesis of Lipid A via the Raetz pathway is essential for the viability and fitness of Gram-negative bacteria, includingChlamydia trachomatis Although nearly all of the enzymes in the Lipid A biosynthetic pathway are highly conserved across Gram-negative bacteria, the cleavage of the pyrophosphate group of UDP-2,3-diacyl-GlcN (UDP-DAGn) to form Lipid X is carried out by two unrelated enzymes: LpXH in beta- and gammaproteobacteria and LpXI in alphaproteobacteria. The intracellular pathogenC. trachomatislacks an ortholog for either of these two enzymes, and yet, it synthesizes Lipid A and eXhibits conservation of genes encoding other Lipid A enzymes. Employing a complementation screen against aC. trachomatisgenomic library using a conditional-lethallpXHmutantEscherichia colistrain, we have identified an open reading frame (Ct461, renamedlpXG) encoding a previously uncharacterized enzyme that complements the UDP-DAGn hydrolase function inE. coliand catalyzes the conversion of UDP-DAGn to Lipid Xin vitro LpXG shows little sequence similarity to either LpXH or LpXI, highlighting LpXG as the founding member of a third class of UDP-DAGn hydrolases. OvereXpression of LpXG results in toXic accumulation of Lipid X and profoundly reduces the infectivity ofC. trachomatis, validating LpXG as the long-sought-after UDP-DAGn pyrophosphatase in this prominent human pathogen. The complementation approach presented here overcomes the lack of suitable genetic tools forC. trachomatisand should be broadly applicable for the functional characterization of other essentialC. trachomatisgenes.IMPORTANCEChlamydia trachomatisis a leading cause of infectious blindness and seXually transmitted disease. Due to the lack of robust genetic tools, the functions of manyChlamydiagenes remain uncharacterized, including the essential gene encoding the UDP-DAGn pyrophosphatase activity for the biosynthesis of Lipid A, the membrane anchor of lipooligosaccharide and the predominant Lipid species of the outer leaflet of the bacterial outer membrane. We designed a complementation screen against theC. trachomatisgenomic library using a conditional-lethal mutant ofE. coliand identified the missing essential gene in the Lipid A biosynthetic pathway, which we designatedlpXG We show that LpXG is a member of the calcineurin-like phosphatases and displays robust UDP-DAGn pyrophosphatase activityin vitro OvereXpression of LpXG inC. trachomatisleads to the accumulation of the predicted Lipid intermediate and reduces bacterial infectivity, validating thein vivofunction of LpXG and highlighting the importance of regulated Lipid A biosynthesis inC. trachomatis.
Jinshi Zhao - One of the best experts on this subject based on the ideXlab platform.
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Structure–Activity Relationship of Sulfonyl Piperazine LpXH Inhibitors Analyzed by an LpXE-Coupled Malachite Green Assay
ACS infectious diseases, 2019Co-Authors: Minhee Lee, Jae Cho, Jinshi Zhao, Seung-hwa Kwak, Myungju Lee, Robert A. Gillespie, Do-yeon Kwon, Hyunji Lee, Hyun-ju ParkAbstract:The UDP-2,3-diacylglucosamine pyrophosphatase LpXH in the Raetz pathway of Lipid A biosynthesis is an essential enzyme in the vast majority of Gram-negative pathogens and an eXcellent novel antibiotic target. The 32P-radioautographic thin-layer chromatography assay has been widely used for analysis of LpXH activity, but it is inconvenient for evaluation of a large number of LpXH inhibitors over an eXtended time period. Here, we report a coupled, nonradioactive LpXH assay that utilizes the recently discovered AquifeX aeolicus Lipid A 1-phosphatase LpXE for quantitative removal of the 1-phosphate from Lipid X, the product of the LpXH catalysis; the released inorganic phosphate is subsequently quantified by the colorimetric malachite green assay, allowing the monitoring of the LpXH catalysis. Using such a coupled enzymatic assay, we report the biochemical characterization of a series of sulfonyl piperazine LpXH inhibitors. Our analysis establishes a preliminary structure–activity relationship for this class ...
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Structure of the essential Haemophilus influenzae UDP-diacylglucosamine pyrophosphohydrolase LpXH in Lipid A biosynthesis
Nature Microbiology, 2016Co-Authors: Jae Cho, Chul-jin Lee, Jinshi Zhao, Hayley E. Young, Pei ZhouAbstract:In most Gram-negative pathogens, the hydrolysis of UDP-2,3-diacylglucosamine to generate Lipid X in Lipid A biosynthesis is catalysed by the membrane-associated enzyme LpXH. We report the crystal structure of LpXH in compleX with its product, Lipid X, unveiling a unique insertion lid above the conserved architecture of calcineurin-like phosphoesterases. This structure reveals elaborate interactions surrounding Lipid X and provides molecular insights into the substrate selectivity, catalysis and inhibition of LpXH. The structure of the LipidA diacylglucosamine pyrophosphohydrolase LpXH bound to its Lipid X substrate is presented.
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Structure of the essential Haemophilus influenzae UDP-diacylglucosamine pyrophosphohydrolase LpXH in Lipid A biosynthesis.
Nature microbiology, 2016Co-Authors: Jae Cho, Chul-jin Lee, Jinshi Zhao, Hayley E. Young, Pei ZhouAbstract:In most Gram-negative pathogens, the hydrolysis of UDP-2,3-diacylglucosamine to generate Lipid X in Lipid A biosynthesis is catalysed by the membrane-associated enzyme LpXH. We report the crystal structure of LpXH in compleX with its product, Lipid X, unveiling a unique insertion lid above the conserved architecture of calcineurin-like phosphoesterases. This structure reveals elaborate interactions surrounding Lipid X and provides molecular insights into the substrate selectivity, catalysis and inhibition of LpXH.
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Discovery of the Elusive UDP-Diacylglucosamine Hydrolase in the Lipid A Biosynthetic Pathway in Chlamydia trachomatis
mBio, 2016Co-Authors: Hayley E. Young, Jinshi Zhao, Jeffrey R. Barker, Ziqiang Guan, Raphael H. Valdivia, Pei ZhouAbstract:Constitutive biosynthesis of Lipid A via the Raetz pathway is essential for the viability and fitness of Gram-negative bacteria, includingChlamydia trachomatis Although nearly all of the enzymes in the Lipid A biosynthetic pathway are highly conserved across Gram-negative bacteria, the cleavage of the pyrophosphate group of UDP-2,3-diacyl-GlcN (UDP-DAGn) to form Lipid X is carried out by two unrelated enzymes: LpXH in beta- and gammaproteobacteria and LpXI in alphaproteobacteria. The intracellular pathogenC. trachomatislacks an ortholog for either of these two enzymes, and yet, it synthesizes Lipid A and eXhibits conservation of genes encoding other Lipid A enzymes. Employing a complementation screen against aC. trachomatisgenomic library using a conditional-lethallpXHmutantEscherichia colistrain, we have identified an open reading frame (Ct461, renamedlpXG) encoding a previously uncharacterized enzyme that complements the UDP-DAGn hydrolase function inE. coliand catalyzes the conversion of UDP-DAGn to Lipid Xin vitro LpXG shows little sequence similarity to either LpXH or LpXI, highlighting LpXG as the founding member of a third class of UDP-DAGn hydrolases. OvereXpression of LpXG results in toXic accumulation of Lipid X and profoundly reduces the infectivity ofC. trachomatis, validating LpXG as the long-sought-after UDP-DAGn pyrophosphatase in this prominent human pathogen. The complementation approach presented here overcomes the lack of suitable genetic tools forC. trachomatisand should be broadly applicable for the functional characterization of other essentialC. trachomatisgenes.IMPORTANCEChlamydia trachomatisis a leading cause of infectious blindness and seXually transmitted disease. Due to the lack of robust genetic tools, the functions of manyChlamydiagenes remain uncharacterized, including the essential gene encoding the UDP-DAGn pyrophosphatase activity for the biosynthesis of Lipid A, the membrane anchor of lipooligosaccharide and the predominant Lipid species of the outer leaflet of the bacterial outer membrane. We designed a complementation screen against theC. trachomatisgenomic library using a conditional-lethal mutant ofE. coliand identified the missing essential gene in the Lipid A biosynthetic pathway, which we designatedlpXG We show that LpXG is a member of the calcineurin-like phosphatases and displays robust UDP-DAGn pyrophosphatase activityin vitro OvereXpression of LpXG inC. trachomatisleads to the accumulation of the predicted Lipid intermediate and reduces bacterial infectivity, validating thein vivofunction of LpXG and highlighting the importance of regulated Lipid A biosynthesis inC. trachomatis.
Jae Cho - One of the best experts on this subject based on the ideXlab platform.
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Structure–Activity Relationship of Sulfonyl Piperazine LpXH Inhibitors Analyzed by an LpXE-Coupled Malachite Green Assay
ACS infectious diseases, 2019Co-Authors: Minhee Lee, Jae Cho, Jinshi Zhao, Seung-hwa Kwak, Myungju Lee, Robert A. Gillespie, Do-yeon Kwon, Hyunji Lee, Hyun-ju ParkAbstract:The UDP-2,3-diacylglucosamine pyrophosphatase LpXH in the Raetz pathway of Lipid A biosynthesis is an essential enzyme in the vast majority of Gram-negative pathogens and an eXcellent novel antibiotic target. The 32P-radioautographic thin-layer chromatography assay has been widely used for analysis of LpXH activity, but it is inconvenient for evaluation of a large number of LpXH inhibitors over an eXtended time period. Here, we report a coupled, nonradioactive LpXH assay that utilizes the recently discovered AquifeX aeolicus Lipid A 1-phosphatase LpXE for quantitative removal of the 1-phosphate from Lipid X, the product of the LpXH catalysis; the released inorganic phosphate is subsequently quantified by the colorimetric malachite green assay, allowing the monitoring of the LpXH catalysis. Using such a coupled enzymatic assay, we report the biochemical characterization of a series of sulfonyl piperazine LpXH inhibitors. Our analysis establishes a preliminary structure–activity relationship for this class ...
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Structure of the essential Haemophilus influenzae UDP-diacylglucosamine pyrophosphohydrolase LpXH in Lipid A biosynthesis
Nature Microbiology, 2016Co-Authors: Jae Cho, Chul-jin Lee, Jinshi Zhao, Hayley E. Young, Pei ZhouAbstract:In most Gram-negative pathogens, the hydrolysis of UDP-2,3-diacylglucosamine to generate Lipid X in Lipid A biosynthesis is catalysed by the membrane-associated enzyme LpXH. We report the crystal structure of LpXH in compleX with its product, Lipid X, unveiling a unique insertion lid above the conserved architecture of calcineurin-like phosphoesterases. This structure reveals elaborate interactions surrounding Lipid X and provides molecular insights into the substrate selectivity, catalysis and inhibition of LpXH. The structure of the LipidA diacylglucosamine pyrophosphohydrolase LpXH bound to its Lipid X substrate is presented.
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Structure of the essential Haemophilus influenzae UDP-diacylglucosamine pyrophosphohydrolase LpXH in Lipid A biosynthesis.
Nature microbiology, 2016Co-Authors: Jae Cho, Chul-jin Lee, Jinshi Zhao, Hayley E. Young, Pei ZhouAbstract:In most Gram-negative pathogens, the hydrolysis of UDP-2,3-diacylglucosamine to generate Lipid X in Lipid A biosynthesis is catalysed by the membrane-associated enzyme LpXH. We report the crystal structure of LpXH in compleX with its product, Lipid X, unveiling a unique insertion lid above the conserved architecture of calcineurin-like phosphoesterases. This structure reveals elaborate interactions surrounding Lipid X and provides molecular insights into the substrate selectivity, catalysis and inhibition of LpXH.
Hayley E. Young - One of the best experts on this subject based on the ideXlab platform.
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Structure of the essential Haemophilus influenzae UDP-diacylglucosamine pyrophosphohydrolase LpXH in Lipid A biosynthesis
Nature Microbiology, 2016Co-Authors: Jae Cho, Chul-jin Lee, Jinshi Zhao, Hayley E. Young, Pei ZhouAbstract:In most Gram-negative pathogens, the hydrolysis of UDP-2,3-diacylglucosamine to generate Lipid X in Lipid A biosynthesis is catalysed by the membrane-associated enzyme LpXH. We report the crystal structure of LpXH in compleX with its product, Lipid X, unveiling a unique insertion lid above the conserved architecture of calcineurin-like phosphoesterases. This structure reveals elaborate interactions surrounding Lipid X and provides molecular insights into the substrate selectivity, catalysis and inhibition of LpXH. The structure of the LipidA diacylglucosamine pyrophosphohydrolase LpXH bound to its Lipid X substrate is presented.
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Structure of the essential Haemophilus influenzae UDP-diacylglucosamine pyrophosphohydrolase LpXH in Lipid A biosynthesis.
Nature microbiology, 2016Co-Authors: Jae Cho, Chul-jin Lee, Jinshi Zhao, Hayley E. Young, Pei ZhouAbstract:In most Gram-negative pathogens, the hydrolysis of UDP-2,3-diacylglucosamine to generate Lipid X in Lipid A biosynthesis is catalysed by the membrane-associated enzyme LpXH. We report the crystal structure of LpXH in compleX with its product, Lipid X, unveiling a unique insertion lid above the conserved architecture of calcineurin-like phosphoesterases. This structure reveals elaborate interactions surrounding Lipid X and provides molecular insights into the substrate selectivity, catalysis and inhibition of LpXH.
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Discovery of the Elusive UDP-Diacylglucosamine Hydrolase in the Lipid A Biosynthetic Pathway in Chlamydia trachomatis
mBio, 2016Co-Authors: Hayley E. Young, Jinshi Zhao, Jeffrey R. Barker, Ziqiang Guan, Raphael H. Valdivia, Pei ZhouAbstract:Constitutive biosynthesis of Lipid A via the Raetz pathway is essential for the viability and fitness of Gram-negative bacteria, includingChlamydia trachomatis Although nearly all of the enzymes in the Lipid A biosynthetic pathway are highly conserved across Gram-negative bacteria, the cleavage of the pyrophosphate group of UDP-2,3-diacyl-GlcN (UDP-DAGn) to form Lipid X is carried out by two unrelated enzymes: LpXH in beta- and gammaproteobacteria and LpXI in alphaproteobacteria. The intracellular pathogenC. trachomatislacks an ortholog for either of these two enzymes, and yet, it synthesizes Lipid A and eXhibits conservation of genes encoding other Lipid A enzymes. Employing a complementation screen against aC. trachomatisgenomic library using a conditional-lethallpXHmutantEscherichia colistrain, we have identified an open reading frame (Ct461, renamedlpXG) encoding a previously uncharacterized enzyme that complements the UDP-DAGn hydrolase function inE. coliand catalyzes the conversion of UDP-DAGn to Lipid Xin vitro LpXG shows little sequence similarity to either LpXH or LpXI, highlighting LpXG as the founding member of a third class of UDP-DAGn hydrolases. OvereXpression of LpXG results in toXic accumulation of Lipid X and profoundly reduces the infectivity ofC. trachomatis, validating LpXG as the long-sought-after UDP-DAGn pyrophosphatase in this prominent human pathogen. The complementation approach presented here overcomes the lack of suitable genetic tools forC. trachomatisand should be broadly applicable for the functional characterization of other essentialC. trachomatisgenes.IMPORTANCEChlamydia trachomatisis a leading cause of infectious blindness and seXually transmitted disease. Due to the lack of robust genetic tools, the functions of manyChlamydiagenes remain uncharacterized, including the essential gene encoding the UDP-DAGn pyrophosphatase activity for the biosynthesis of Lipid A, the membrane anchor of lipooligosaccharide and the predominant Lipid species of the outer leaflet of the bacterial outer membrane. We designed a complementation screen against theC. trachomatisgenomic library using a conditional-lethal mutant ofE. coliand identified the missing essential gene in the Lipid A biosynthetic pathway, which we designatedlpXG We show that LpXG is a member of the calcineurin-like phosphatases and displays robust UDP-DAGn pyrophosphatase activityin vitro OvereXpression of LpXG inC. trachomatisleads to the accumulation of the predicted Lipid intermediate and reduces bacterial infectivity, validating thein vivofunction of LpXG and highlighting the importance of regulated Lipid A biosynthesis inC. trachomatis.
Min Yao - One of the best experts on this subject based on the ideXlab platform.
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Crystal structures of the UDP-diacylglucosamine pyrophosphohydrase LpXH from Pseudomonas aeruginosa.
Scientific reports, 2016Co-Authors: Chiaki Okada, Hiroko Wakabayashi, Momoko Kobayashi, Akira Shinoda, Isao Tanaka, Min YaoAbstract:Lipid A (also known as endotoXin) is the hydrophobic portion of lipopolysaccharides. It is an essential membrane component required for the viability of gram-negative bacteria. The enzymes involved in its biosynthesis are attractive targets for the development of novel antibiotics. LpXH catalyzes the fourth step of the Lipid A biosynthesis pathway and cleaves the pyrophosphate bond of UDP-2,3-diacylglucosamine to yield 2,3-diacylglucosamine 1-phosphate (Lipid X) and UMP. Here we present the structures of LpXH from Pseudomonas aeruginosa (PaLpXH). PaLpXH consists of two domains: a catalytic domain that is homologous to the metallophosphoesterases and a helical insertion domain. Lipid X was captured in the crevice between these two domains, with its phosphate group facing the dinuclear metal (Mn2+) center and two acyl chains buried in the hydrophobic cavity. The structures reveal that a large conformational change occurs at the Lipid X binding site surface upon the binding/release of the product molecule. Based on these observations, we propose a novel model for Lipid X embedding, which involves the scissor-like movement of heliX α6, resulting in the release of Lipid X into the Lipid bilayer.