Cloning, expression and characterization of a 46.5-kDa metallopeptidase from Bacillus halodurans H4 sharing properties with the pitrilysin family

Dabonne, Soumaila; Moallic, Claire; Sine, Jean‐Pierre; Niamké, Sebastien; Dion, Michel; Colas, Bernard

doi:10.1016/j.bbagen.2005.03.016

Cited by 17 publications

(16 citation statements)

References 37 publications

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“…Of these, two cleavage sites, Tyr16-Leu17 and Phe25-Tyr26, are also targeted by human insulysin, 27,28 E. coli pitrilysin, 29 and B. halodurans ppBH4. 22 Phe24-Phe25 cleavage is also observed in human insulysin activity. 27,28 Cleavage sites within dynorphin A(1-13) were also determined (Table 3).…”

Section: Enzyme Characteristics Of Strain A1 M16b Peptidasementioning

confidence: 77%

“…Of these, only Phe4-Leu5 is commonly cleaved by another M16B peptidase, ppBH4. 22 To the best of our knowledge, SPH2681/SPH2682 is the first example of a bacterial heterosubunit-type M16B peptidase. Since the initiation of our experiments investigating this peptidase, 30 other groups have predicted the possibility of heterodimer formation in bacteria based on genetic organization …”

Section: Enzyme Characteristics Of Strain A1 M16b Peptidasementioning

confidence: 99%

“…21 Among the M16 enzymes lacking the glycine-rich loop, an R/Y pair is conserved in the C-terminal domain. Until recently, bacterial M16B enzymes have been suggested to be active in monomeric form based on the enzymology of Bacillus halodurans peptidase ppBH4 22 and Rickettsia prowazekii peptidase RPP. 23 On the other hand, the ppBH4-homologous BHP from another strain of B. halodurans has been recently reported to form an active homodimer.…”

Section: Introductionmentioning

confidence: 99%

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Heterosubunit Composition and Crystal Structures of a Novel Bacterial M16B Metallopeptidase

Maruyama¹,

Chuma²,

Mikami³

et al. 2011

Journal of Molecular Biology

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Section: Enzyme Characteristics Of Strain A1 M16b Peptidasementioning

confidence: 77%

Section: Enzyme Characteristics Of Strain A1 M16b Peptidasementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Heterosubunit Composition and Crystal Structures of a Novel Bacterial M16B Metallopeptidase

Maruyama¹,

Chuma²,

Mikami³

et al. 2011

Journal of Molecular Biology

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“…DTT is a strong reducing agent. Since there are four cysteine residues in KPN_02808, the inhibition of the enzymatic activity by DTT might be due to the disruption of disulfide bonds that stabilize the protein [8]. The enzyme was not inhibited or sensitized by PMSF, suggesting that the KPN_02809 protein was not a serine protease.…”

Section: Resultsmentioning

confidence: 99%

“…Metalloprotease, the most diverse of the six main types of proteases, has drawn much of our interest as it plays an important role in host-pathogen interactions by promoting enteropathogenicity, vascular permeability, host tissue damage and cytotoxicity [7]. Metalloproteases expressed by pathogens such as Legionella pneumophila , Vibrio cholerae and Plasmodium vivax involve in pathogenesis of the disease by degrading a wide range of host molecules [8–10]. …”

Section: Introductionmentioning

confidence: 99%

Structural Modeling and Biochemical Characterization of Recombinant KPN_02809, a Zinc-Dependent Metalloprotease from Klebsiella pneumoniae MGH 78578

Wong

Choi

Kuan

et al. 2012

IJMS

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Klebsiella pneumoniae is a Gram-negative, cylindrical rod shaped opportunistic pathogen that is found in the environment as well as existing as a normal flora in mammalian mucosal surfaces such as the mouth, skin, and intestines. Clinically it is the most important member of the family of Enterobacteriaceae that causes neonatal sepsis and nosocomial infections. In this work, a combination of protein sequence analysis, structural modeling and molecular docking simulation approaches were employed to provide an understanding of the possible functions and characteristics of a hypothetical protein (KPN_02809) from K. pneumoniae MGH 78578. The computational analyses showed that this protein was a metalloprotease with zinc binding motif, HEXXH. To verify this result, a ypfJ gene which encodes for this hypothetical protein was cloned from K. pneumoniae MGH 78578 and the protein was overexpressed in Escherichia coli BL21 (DE3). The purified protein was about 32 kDa and showed maximum protease activity at 30 °C and pH 8.0. The enzyme activity was inhibited by metalloprotease inhibitors such as EDTA, 1,10-phenanthroline and reducing agent, 1,4-dithiothreitol (DTT). Each molecule of KPN_02809 protein was also shown to bind one zinc ion. Hence, for the first time, we experimentally confirmed that KPN_02809 is an active enzyme with zinc metalloprotease activity.

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The burden of fatigue and quality of life in myeloproliferative disorders (MPDs)

Mesa

Niblack

Wadleigh

et al. 2006

Cancer

355

243

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Cloning, expression and characterization of a 46.5-kDa metallopeptidase from Bacillus halodurans H4 sharing properties with the pitrilysin family

Cited by 17 publications

References 37 publications

Heterosubunit Composition and Crystal Structures of a Novel Bacterial M16B Metallopeptidase

Heterosubunit Composition and Crystal Structures of a Novel Bacterial M16B Metallopeptidase

Structural Modeling and Biochemical Characterization of Recombinant KPN_02809, a Zinc-Dependent Metalloprotease from Klebsiella pneumoniae MGH 78578

The burden of fatigue and quality of life in myeloproliferative disorders (MPDs)

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