The leucine aminopeptidase of Staphylococcus aureus is secreted and contributes to biofilm formation

Arun Kumar Singh; Rochika Singh; Dhanendra Tomar; Chirayu D. Pandya; Rajesh Singh

doi:10.1016/j.ijid.2012.01.009

The leucine aminopeptidase of Staphylococcus aureus is secreted and contributes to biofilm formation

Arun Kumar Singh, Rochika Singh, Dhanendra Tomar, Chirayu D. Pandya, Rajesh Singh

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

Background: Staphylococcus aureus has emerged as a major drug-resistant pathogen in hospital- and community-acquired infections. Leucine aminopeptidase (LAP) is known to be essential for survival of the bacteria; however the LAP of S. aureus has not been extensively characterized. In this study, we report a detailed characterization of the S. aureus LAP. Methods: LAP from S. aureus was cloned, purified, and further biochemically characterized. The expression of LAP was analyzed by Western blotting. Growth and biofilm formation were analyzed spectrophotometrically. Results: LAP was cloned from S. aureus and expressed as a 55 kDa protein, whereas the molecular weight of the native protein is approximately 600 kDa. LAP showed amidolytic activity against l-leucine p-nitroanilide. Optimal activity was observed at pH 8.5 and 37°C with a V_max of 2500μmol/min/mg protein. LAP enzymatic activity was inhibited by ion chelators and enhanced by divalent metal ions, specifically Ni. LAP is secreted by laboratory as well as clinical strains. Bestatin, an inhibitor of LAP, inhibits S. aureus growth and biofilm formation. Conclusions: To our knowledge, this is the first detailed characterization of LAP from S. aureus and suggests its importance in survival and pathogenesis.

Original language	English
Pages (from-to)	e375-e381
Journal	International Journal of Infectious Diseases
Volume	16
Issue number	5
DOIs	https://doi.org/10.1016/j.ijid.2012.01.009
State	Published - May 2012

Bibliographical note

Funding Information:
We are thankful to Prof. M.M. Godbole (S.G. Postgraduate Institute of Medical Sciences, Lucknow, India) for help in generating the polyclonal antibody, and Dr Preena Bhalla (Staphylococcal Phage Typing Center, Department of Microbiology, Maulana Azad Medical College, New Delhi, India) for providing clinical strains of S. aureus . Dhanendra Tomar received a fellowship from the Council of Scientific and Industrial Research (CSIR), New Delhi, India. We are thankful to Dr Anton M. Jetten, Chief, Lab Cell Biology, National Institute of Environmental Health Sciences/National Institutes of Health, North Carolina, USA, for a critical reading of the manuscript. We also acknowledge The Puri Foundation for Education in India for financial support to initiate the work and partial financial support from the Indian Council of Medical Research (ICMR), New Delhi, India.

Keywords

Bestatin
Biofilm
Leucine aminopeptidase
Staphylococcus aureus

ASJC Scopus subject areas

Microbiology (medical)
Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.ijid.2012.01.009

Cite this

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title = "The leucine aminopeptidase of Staphylococcus aureus is secreted and contributes to biofilm formation",

abstract = "Background: Staphylococcus aureus has emerged as a major drug-resistant pathogen in hospital- and community-acquired infections. Leucine aminopeptidase (LAP) is known to be essential for survival of the bacteria; however the LAP of S. aureus has not been extensively characterized. In this study, we report a detailed characterization of the S. aureus LAP. Methods: LAP from S. aureus was cloned, purified, and further biochemically characterized. The expression of LAP was analyzed by Western blotting. Growth and biofilm formation were analyzed spectrophotometrically. Results: LAP was cloned from S. aureus and expressed as a 55 kDa protein, whereas the molecular weight of the native protein is approximately 600 kDa. LAP showed amidolytic activity against l-leucine p-nitroanilide. Optimal activity was observed at pH 8.5 and 37°C with a Vmax of 2500μmol/min/mg protein. LAP enzymatic activity was inhibited by ion chelators and enhanced by divalent metal ions, specifically Ni. LAP is secreted by laboratory as well as clinical strains. Bestatin, an inhibitor of LAP, inhibits S. aureus growth and biofilm formation. Conclusions: To our knowledge, this is the first detailed characterization of LAP from S. aureus and suggests its importance in survival and pathogenesis.",

keywords = "Bestatin, Biofilm, Leucine aminopeptidase, Staphylococcus aureus",

author = "Singh, {Arun Kumar} and Rochika Singh and Dhanendra Tomar and Pandya, {Chirayu D.} and Rajesh Singh",

note = "Funding Information: We are thankful to Prof. M.M. Godbole (S.G. Postgraduate Institute of Medical Sciences, Lucknow, India) for help in generating the polyclonal antibody, and Dr Preena Bhalla (Staphylococcal Phage Typing Center, Department of Microbiology, Maulana Azad Medical College, New Delhi, India) for providing clinical strains of S. aureus . Dhanendra Tomar received a fellowship from the Council of Scientific and Industrial Research (CSIR), New Delhi, India. We are thankful to Dr Anton M. Jetten, Chief, Lab Cell Biology, National Institute of Environmental Health Sciences/National Institutes of Health, North Carolina, USA, for a critical reading of the manuscript. We also acknowledge The Puri Foundation for Education in India for financial support to initiate the work and partial financial support from the Indian Council of Medical Research (ICMR), New Delhi, India. ",

year = "2012",

month = may,

doi = "10.1016/j.ijid.2012.01.009",

language = "English",

volume = "16",

pages = "e375--e381",

journal = "International Journal of Infectious Diseases",

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T1 - The leucine aminopeptidase of Staphylococcus aureus is secreted and contributes to biofilm formation

AU - Singh, Arun Kumar

AU - Singh, Rochika

AU - Tomar, Dhanendra

AU - Pandya, Chirayu D.

AU - Singh, Rajesh

N1 - Funding Information: We are thankful to Prof. M.M. Godbole (S.G. Postgraduate Institute of Medical Sciences, Lucknow, India) for help in generating the polyclonal antibody, and Dr Preena Bhalla (Staphylococcal Phage Typing Center, Department of Microbiology, Maulana Azad Medical College, New Delhi, India) for providing clinical strains of S. aureus . Dhanendra Tomar received a fellowship from the Council of Scientific and Industrial Research (CSIR), New Delhi, India. We are thankful to Dr Anton M. Jetten, Chief, Lab Cell Biology, National Institute of Environmental Health Sciences/National Institutes of Health, North Carolina, USA, for a critical reading of the manuscript. We also acknowledge The Puri Foundation for Education in India for financial support to initiate the work and partial financial support from the Indian Council of Medical Research (ICMR), New Delhi, India.

PY - 2012/5

Y1 - 2012/5

N2 - Background: Staphylococcus aureus has emerged as a major drug-resistant pathogen in hospital- and community-acquired infections. Leucine aminopeptidase (LAP) is known to be essential for survival of the bacteria; however the LAP of S. aureus has not been extensively characterized. In this study, we report a detailed characterization of the S. aureus LAP. Methods: LAP from S. aureus was cloned, purified, and further biochemically characterized. The expression of LAP was analyzed by Western blotting. Growth and biofilm formation were analyzed spectrophotometrically. Results: LAP was cloned from S. aureus and expressed as a 55 kDa protein, whereas the molecular weight of the native protein is approximately 600 kDa. LAP showed amidolytic activity against l-leucine p-nitroanilide. Optimal activity was observed at pH 8.5 and 37°C with a Vmax of 2500μmol/min/mg protein. LAP enzymatic activity was inhibited by ion chelators and enhanced by divalent metal ions, specifically Ni. LAP is secreted by laboratory as well as clinical strains. Bestatin, an inhibitor of LAP, inhibits S. aureus growth and biofilm formation. Conclusions: To our knowledge, this is the first detailed characterization of LAP from S. aureus and suggests its importance in survival and pathogenesis.

AB - Background: Staphylococcus aureus has emerged as a major drug-resistant pathogen in hospital- and community-acquired infections. Leucine aminopeptidase (LAP) is known to be essential for survival of the bacteria; however the LAP of S. aureus has not been extensively characterized. In this study, we report a detailed characterization of the S. aureus LAP. Methods: LAP from S. aureus was cloned, purified, and further biochemically characterized. The expression of LAP was analyzed by Western blotting. Growth and biofilm formation were analyzed spectrophotometrically. Results: LAP was cloned from S. aureus and expressed as a 55 kDa protein, whereas the molecular weight of the native protein is approximately 600 kDa. LAP showed amidolytic activity against l-leucine p-nitroanilide. Optimal activity was observed at pH 8.5 and 37°C with a Vmax of 2500μmol/min/mg protein. LAP enzymatic activity was inhibited by ion chelators and enhanced by divalent metal ions, specifically Ni. LAP is secreted by laboratory as well as clinical strains. Bestatin, an inhibitor of LAP, inhibits S. aureus growth and biofilm formation. Conclusions: To our knowledge, this is the first detailed characterization of LAP from S. aureus and suggests its importance in survival and pathogenesis.

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KW - Leucine aminopeptidase

KW - Staphylococcus aureus

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SN - 1201-9712

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SP - e375-e381

JO - International Journal of Infectious Diseases

JF - International Journal of Infectious Diseases

IS - 5

ER -

The leucine aminopeptidase of Staphylococcus aureus is secreted and contributes to biofilm formation

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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