Mycobacterial lipolytic enzymes: A gold mine for tuberculosis research

Dedieu, Luc; Serveau-Avesque, Carole; Kremer, Laurent; Canaan, Stéphane

doi:10.1016/j.biochi.2012.07.008

Cited by 58 publications

(48 citation statements)

References 51 publications

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“…Schué et al (28) isolated two secreted cutinases-like proteins: Rv1984c, which preferentially hydrolyzed medium-chain carboxylic esters and monoacylglycerols, and Rv3452, which behaved like a phospholipase A2. The other lipases/cutinases are most likely cell-wall-associated and surface-exposed (29). Concerning the subsequent steps in recycling of phospholipids, in the M. tuberculosis genome, Rv0317c (glpQ2) and Rv3842c (glpQ1) are annotated as glycerophosphoryl diester phosphodiesterases that may degrade polar heads generating glycerol 3-phosphate and the corresponding alcohols.…”

Section: Resultsmentioning

confidence: 99%

Discovery of a glycerol 3-phosphate phosphatase reveals glycerophospholipid polar head recycling in Mycobacterium tuberculosis

Larrouy-Maumus

Biswas

Hunt

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Functional assignment of enzymes encoded by the Mycobacterium tuberculosis genome is largely incomplete despite recent advances in genomics and bioinformatics. Here, we applied an activitybased metabolomic profiling method to assign function to a unique phosphatase, Rv1692. In contrast to its annotation as a nucleotide phosphatase, metabolomic profiling and kinetic characterization indicate that Rv1692 is a D,L-glycerol 3-phosphate phosphatase. Crystal structures of Rv1692 reveal a unique architecture, a fusion of a predicted haloacid dehalogenase fold with a previously unidentified GCN5-related N-acetyltransferase region.Although not directly involved in acetyl transfer, or regulation of enzymatic activity in vitro, this GCN5-related N-acetyltransferase region is critical for the solubility of the phosphatase. Structural and biochemical analysis shows that the active site features are adapted for recognition of small polyol phosphates, and not nucleotide substrates. Functional assignment and metabolomic studies of M. tuberculosis lacking rv1692 demonstrate that Rv1692 is the final enzyme involved in glycerophospholipid recycling/catabolism, a pathway not previously described in M. tuberculosis.haloacid dehalogenase superfamily | enzyme function | pathway discovery E ach year, 1.4 million people succumb to tuberculosis, making Mycobacterium tuberculosis the deadliest bacterium affecting mankind (1). In addition, the dissemination of strains resistant to several antibiotics underscores the need for better understanding of this pathogen and for the development of novel vaccines and therapeutics (2, 3). Our approach to elucidating the unique pervasiveness of M. tuberculosis is through comprehensive discovery and characterization of metabolic pathways, as metabolism underlies survival of the bacteria both inside and outside the host and can contribute to phenotypic and genetic drug resistance.The M. tuberculosis genome encodes for 4,043 genes, of which 3,933 encode proteins (4, 5). Many genes with essential functions, such as DNA replication, protein and RNA synthesis, and cell-division, have close homologs in other bacteria, and their functions are annotated largely on the basis of analysis of their counterparts. However, functions of at least one-third of the genes are unknown or putative (4). In particular, little is known about genes that are not conserved or conditionally important. Characterization of such genes presents a daunting task. M. tuberculosis is thought to be subjected to a myriad of conditions during its life cycle in the host, such as low pH, reactive oxygen and nitrogen species, and so on (6, 7). Understanding how M. tuberculosis adapts to and even thrives in such diverse environments is critical to our understanding of the pathology itself and for the discovery of novel therapeutics to treat tuberculosis.We applied an innovative, activity-based metabolomic profiling (ABMP) approach to assign function to orphan (without a priori known substrates/products) mycobacterial enzymes and to discover unk...

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Section: Resultsmentioning

confidence: 99%

Discovery of a glycerol 3-phosphate phosphatase reveals glycerophospholipid polar head recycling in Mycobacterium tuberculosis

Larrouy-Maumus

Biswas

Hunt

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…We next extended this approach to the study of LipH where natural substrates are less well characterized (39,45). Threedimensional models of LipH (Fig.…”

Section: Figmentioning

confidence: 99%

Targeting Lipid Esterases in Mycobacteria Grown Under Different Physiological Conditions Using Activity-based Profiling with Tetrahydrolipstatin (THL)

Ravindran

Rao

Cheng

et al. 2014

Molecular & Cellular Proteomics

104

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Tetrahydrolipstatin (THL) is bactericidal but its precise target spectrum is poorly characterized. Here, we used a THL analog and activity-based protein profiling to identify target proteins after enrichment from whole cell lysates of Mycobacterium bovis Bacillus Calmette-Gué rin cultured under replicating and non-replicating conditions. THL targets ␣/␤-hydrolases, including many lipid esterases (LipD, G, H, I, M, N, O, V, W, and TesA). Target protein concentrations and total esterase activity correlated inversely with cellular triacylglycerol upon entry into and exit from non-replicating conditions. Cellular overexpression of lipH and tesA led to decreased THL susceptibility thus providing functional validation. Our results define the target spectrum of THL in a biological species with particularly diverse lipid metabolic pathways. We furthermore derive a conceptual approach that demonstrates the use of such THL probes for the characterization of substrate recognition by lipases and related enzymes. Molecular & Cellular

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“…tuberculosis enters the host by inhalation, and once in the lungs is phagocytosed by macrophages, which may lead to its elimination or to a persistent infection with the formation of granulomas having foamy macrophages containing bacteria (267). M. tuberculosis accumulates inclusions of host cell membrane-derived lipids and remains dormant, or under host immune depression can become active, replicate, and spread into the lung and other tissues (267). The genome of most M. tuberculosis strains encodes various lipases and up to four PLCs, encoded by plcA/rv2351c, plcB/rv2350c, plcC/rv2349c, and plcD/rv1755c, two of which have SMase C activity (267,268).…”

Section: Fig 11mentioning

confidence: 99%

“…M. tuberculosis accumulates inclusions of host cell membrane-derived lipids and remains dormant, or under host immune depression can become active, replicate, and spread into the lung and other tissues (267). The genome of most M. tuberculosis strains encodes various lipases and up to four PLCs, encoded by plcA/rv2351c, plcB/rv2350c, plcC/rv2349c, and plcD/rv1755c, two of which have SMase C activity (267,268). These PLCs are strongly upregulated during the first 24 h of macrophage infection and are cytotoxic to mouse macrophages (268,269).…”

Section: Fig 11mentioning

confidence: 99%

“…PlcA and PlcB have been experimentally shown to be transferred through the cytoplasmic membrane by means of the Tat system (270). Phospholipid hydrolysis by M. tuberculosis PLCs releases DAG, which may be hydrolyzed by lipases, releasing fatty acids that serve either as a carbon source or as building blocks for cell wall lipid synthesis in chronically infected lungs (267). Although a quadruple mutant, M. tuberculosis plcABCD, was reported to be attenuated in its growth kinetics during the late phase of pulmonary infection in mice (268), in a recent study no differences were found between wild-type and mutant strains lacking PLCs in their phagosomal escape, growth abilities within macrophages, or in a mouse infection model (271).…”

Section: Fig 11mentioning

confidence: 99%

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Bacterial Sphingomyelinases and Phospholipases as Virulence Factors

Flores-Dı́az

Monturiol-Gross

Naylor

et al. 2016

Microbiol Mol Biol Rev

165

143

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SUMMARYBacterial sphingomyelinases and phospholipases are a heterogeneous group of esterases which are usually surface associated or secreted by a wide variety of Gram-positive and Gram-negative bacteria. These enzymes hydrolyze sphingomyelin and glycerophospholipids, respectively, generating products identical to the ones produced by eukaryotic enzymes which play crucial roles in distinct physiological processes, including membrane dynamics, cellular signaling, migration, growth, and death. Several bacterial sphingomyelinases and phospholipases are essential for virulence of extracellular, facultative, or obligate intracellular pathogens, as these enzymes contribute to phagosomal escape or phagosomal maturation avoidance, favoring tissue colonization, infection establishment and progression, or immune response evasion. This work presents a classification proposal for bacterial sphingomyelinases and phospholipases that considers not only their enzymatic activities but also their structural aspects. An overview of the main physiopathological activities is provided for each enzyme type, as are examples in which inactivation of a sphingomyelinase- or a phospholipase-encoding gene impairs the virulence of a pathogen. The identification of sphingomyelinases and phospholipases important for bacterial pathogenesis and the development of inhibitors for these enzymes could generate candidate vaccines and therapeutic agents, which will diminish the impacts of the associated human and animal diseases.

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Mycobacterial lipolytic enzymes: A gold mine for tuberculosis research

Cited by 58 publications

References 51 publications

Discovery of a glycerol 3-phosphate phosphatase reveals glycerophospholipid polar head recycling in Mycobacterium tuberculosis

Discovery of a glycerol 3-phosphate phosphatase reveals glycerophospholipid polar head recycling in Mycobacterium tuberculosis

Targeting Lipid Esterases in Mycobacteria Grown Under Different Physiological Conditions Using Activity-based Profiling with Tetrahydrolipstatin (THL)

Bacterial Sphingomyelinases and Phospholipases as Virulence Factors

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