Crystal Structure of the LasA Virulence Factor from Pseudomonas aeruginosa: Substrate Specificity and Mechanism of M23 Metallopeptidases

Spencer, James; Murphy, Loretta M.; Conners, R.; Sessions, Richard B.; Gamblin, S.J.

doi:10.1016/j.jmb.2009.12.021

Cited by 59 publications

(79 citation statements)

References 96 publications

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“…In addition, the structure includes three disulfide bonds, Cys 7 Fig. 2A) (24,26,28). These three classical M23 family members all specifically target the pentaglycine cross-links present in S. aureus peptidoglycan.…”

Section: Resultsmentioning

confidence: 99%

“…These three classical M23 family members all specifically target the pentaglycine cross-links present in S. aureus peptidoglycan. Furthermore, LasA can cleave a wider range of glycine-rich peptides with aromatic or branched amino acids at the P1Ј position such as Pro-Gly-Gly2Tyr-Gly and Leu-Gly-Gly2Ala-Gly (2denotes the cleavage site) (24,26,28,33). A superimposition of the structures of LECT2 and LytM revealed that the central ␤-sheet of LECT2 was similar to that of LytM, which consists of six antiparallel ␤-strands with a conserved topology, but loops 1-4 in the two structures showed significant differences in the lengths and conformations (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…They are divided into two subclasses based on sequence homology: the M23A family, which includes the ␤-lytic endopeptidase from Lysobacter enzymogenes and LasA from Pseudomonas aeruginosa, and the M23B family, which consists of the majority of the family members, including LytM from Staphylococcus aureus (22,24). The M23A endopeptidases possess broader substrate specificities than those of the M23B endopeptidases (22,24). Proteins of the M23 family are distinguished by conserved Zn(II)-binding res-idues that occur in the HX n D (HXXXD for M23B) and HXH motifs (24 -28).…”

Section: Leukocyte Cell-derived Chemotaxin 2 (Lect2)mentioning

confidence: 99%

“…These residues allow Zn(II) to polarize the substrate carbonyl oxygen to increase the susceptibility to nucleophilic attack. An active site histidine that does not coordinate Zn(II) abstracts a proton from Zn(II)-bound water and donates it to the departing amide nitrogen of the substrate, resulting in cleavage of the peptide bond (24,29).…”

Section: Leukocyte Cell-derived Chemotaxin 2 (Lect2)mentioning

confidence: 99%

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Crystal Structure of Human Leukocyte Cell-derived Chemotaxin 2 (LECT2) Reveals a Mechanistic Basis of Functional Evolution in a Mammalian Protein with an M23 Metalloendopeptidase Fold

Zheng

Miyakawa

Sawano

et al. 2016

Journal of Biological Chemistry

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Human leukocyte cell-derived chemotaxin 2 (LECT2), which is predominantly expressed in the liver, is a multifunctional protein. LECT2 is becoming a potential therapeutic target for several diseases of worldwide concern such as rheumatoid arthritis, hepatocellular carcinoma, and obesity. Here, we present the crystal structure of LECT2, the first mammalian protein whose structure contains an M23 metalloendopeptidase fold. The LECT2 structure adopts a conserved Zn(II) coordination configuration but lacks a proposed catalytic histidine residue, and its potential substratebinding groove is blocked in the vicinity of the Zn(II)-binding site by an additional intrachain loop at the N terminus. Consistent with these structural features, LECT2 was found to be catalytically inactive as a metalloendopeptidase against various types of peptide sequences, including pentaglycine. In addition, a surface plasmon resonance analysis demonstrated that LECT2 bound to the c-Met receptor with micromolar affinity. These results indicate that LECT2 likely plays its critical roles by acting as a ligand for the corresponding protein receptors rather than as an enzymatically active peptidase. The intrachain loop together with the pseudoactive site groove in LECT2 structure may be specific for interactions between LECT2 and receptors. Our study reveals a mechanistic basis for the functional evolution of a mammalian protein with an M23 metalloendopeptidase fold and potentially broadens the implications for the biological importance of noncatalytic peptidases in the M23 family.Leukocyte cell-derived chemotaxin 2 (LECT2) 2 is a secretory protein originally identified as a chemotactic factor for neutrophils (1). LECT2 is predominantly expressed in the liver and is a direct target gene of Wnt/␤-catenin signaling (2-6). Accumulating evidence shows that mammalian LECT2 is a multifunctional protein that is closely associated with several diseases of worldwide concern, including hepatitis (7), rheumatoid arthritis (8 -10), hepatocellular carcinoma (HCC) (11, 12), obesity (13,14), and renal and hepatic amyloidosis (15-17). It has been reported that concanavalin A-induced hepatitis and collagen antibody-induced arthritis were suppressed by LECT2 (7,8). In addition, LECT2 expression inhibited the migration and invasion of human HCC cells in vitro and was negatively correlated with vascular invasion and tumor recurrence in HCC patients (11). In contrast, serum LECT2 levels were positively correlated with the severity of obesity and fatty liver in humans, and overproduction of LECT2 caused the development of obesity-associated insulin resistance (13,14). These findings suggest that LECT2 may be a candidate prognostic marker and a potential therapeutic target for these diseases. Despite the importance of LECT2 functions, the underlying mechanisms remain largely unclear.The mature human LECT2 is a basic protein consisting of 133 amino acids. Sequence similarity searches using BLAST have shown that LECT2 has a putative peptidase-M23 (PF01551) domain located ...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Leukocyte Cell-derived Chemotaxin 2 (Lect2)mentioning

confidence: 99%

Section: Leukocyte Cell-derived Chemotaxin 2 (Lect2)mentioning

confidence: 99%

See 2 more Smart Citations

Crystal Structure of Human Leukocyte Cell-derived Chemotaxin 2 (LECT2) Reveals a Mechanistic Basis of Functional Evolution in a Mammalian Protein with an M23 Metalloendopeptidase Fold

Zheng

Miyakawa

Sawano

et al. 2016

Journal of Biological Chemistry

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“…ADAM17 activation also mediates release of soluble syndecan-1 and -4 into the bronchoalveolar fluid of mice, as evidenced by the fact that both constitutive and induced syndecan shedding could be prevented by inhibiting ADAM17 (37). The pseudomonas virulence factor LasA, an M23 metallopeptidase related to autolytic glycylglycine endopeptidases, is known for stimulating ectodomain shedding of syndecan-1 (38,39). Collectively, these observations establish that shedding of syndecans is mediated by a number of factors, including MMP-9, MMP-7, MT1-MMP, ADMTS1, ADAM17, and LasA.…”

Section: Syndecan Shedding Enzymesmentioning

confidence: 99%

Shedding; towards a new paradigm of syndecan function in cancer

Choi

Lee²,

Choi³

et al. 2010

BMB Reports

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Exploration of Phytoconstituents from Mussaenda roxburghii and Studies of Their Antibiofilm Effect

Ghosh

Das

Sarkar

et al. 2017

Chemistry & Biodiversity

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In the context of ethno botanical importance with no phytochemical investigations, Mussaenda roxburghii have been investigated to explore it's phytoconstituents and studies of their antibiofilm activity. Four compounds have been isolated from the aerial parts of this plant and were characterized as 2α,3β,19α,23-tetrahydroxyurs-12-en-28-oic acid (1), β-sitosterol glucoside (4), lupeol palmitate (5), and myoinositol (6). All these compounds were tested for antibacterial and antibiofilm activity against Pseudomonas aeruginosa. Compound 1 exhibited three times more antibiofilm activity with minimum inhibitory concentration (MIC) at 0.74 mm compared to that of streptomycin. Molecular docking studies exhibited a very high binding affinity of 1 with P. aeruginosa quorum sensing proteins and motility associated proteins viz. LasR and PilB, PilY1, PilT, respectively. Compound 1 was also found to be non-cytotoxic against sheep RBC and murine peritoneal macrophages at selected sub-MIC doses.

show abstract

Crystal Structure of the LasA Virulence Factor from Pseudomonas aeruginosa: Substrate Specificity and Mechanism of M23 Metallopeptidases

Cited by 59 publications

References 96 publications

Crystal Structure of Human Leukocyte Cell-derived Chemotaxin 2 (LECT2) Reveals a Mechanistic Basis of Functional Evolution in a Mammalian Protein with an M23 Metalloendopeptidase Fold

Crystal Structure of Human Leukocyte Cell-derived Chemotaxin 2 (LECT2) Reveals a Mechanistic Basis of Functional Evolution in a Mammalian Protein with an M23 Metalloendopeptidase Fold

Shedding; towards a new paradigm of syndecan function in cancer

Exploration of Phytoconstituents from Mussaenda roxburghii and Studies of Their Antibiofilm Effect

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