Solution structure and dynamics of C-terminal regulatory domain of Vibrio vulnificus extracellular metalloprotease

Ji Hye Yun, Heeyoun Kim, Jung Eun Park, Jung Sup Lee, Weon Tae Lee

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

An extracellular metalloprotease (vEP) secreted by Vibrio vulnificus ATCC29307 is a 45-kDa proteolytic enzyme that has prothrombin activation and fibrinolytic activities during bacterial infection. The action of vEP could result in clotting that could serve to protect the bacteria from the host defense machinery. Very recently, we showed that the C-terminal propeptide (C-ter100), which is unique to vEP, is involved in regulation of vEP activity. To understand the structural basis of this function of vEP C-ter100, we have determined the solution structure and backbone dynamics using multidimensional nuclear magnetic resonance spectroscopy. The solution structure shows that vEP C-ter100 is composed of eight anti-parallel β-strands with a unique fold that has a compact β-barrel formation which stabilized by hydrophobic and hydrogen bonding networks. Protein dynamics shows that the overall structure, including loops, is very rigid and stabilized. By structural database analysis, we found that vEP C-ter100 shares its topology with that of the collagen-binding domain of collagenase, despite low sequence homology between the two domains. Fluorescence assay reveals that vEP C-ter100 interacts strongly with iron (Fe 3+ ). These findings suggest that vEP protease might recruit substrate molecules, such as collagen, by binding at C-ter100 and that vEP participates in iron uptake from iron-withholding proteins of the host cell during infection.

Original languageEnglish
Pages (from-to)541-546
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume430
Issue number2
DOIs
Publication statusPublished - 2013 Jan 11

Fingerprint

Vibrio vulnificus
Metalloproteases
Iron
Peptide Hydrolases
Collagen
Prothrombin
Collagenases
Hydrogen Bonding
Sequence Homology
Bacterial Infections
Nuclear magnetic resonance spectroscopy
Machinery
Assays
Bacteria
Hydrogen bonds
Proteins
Magnetic Resonance Spectroscopy
Fluorescence
Chemical activation
Topology

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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title = "Solution structure and dynamics of C-terminal regulatory domain of Vibrio vulnificus extracellular metalloprotease",
abstract = "An extracellular metalloprotease (vEP) secreted by Vibrio vulnificus ATCC29307 is a 45-kDa proteolytic enzyme that has prothrombin activation and fibrinolytic activities during bacterial infection. The action of vEP could result in clotting that could serve to protect the bacteria from the host defense machinery. Very recently, we showed that the C-terminal propeptide (C-ter100), which is unique to vEP, is involved in regulation of vEP activity. To understand the structural basis of this function of vEP C-ter100, we have determined the solution structure and backbone dynamics using multidimensional nuclear magnetic resonance spectroscopy. The solution structure shows that vEP C-ter100 is composed of eight anti-parallel β-strands with a unique fold that has a compact β-barrel formation which stabilized by hydrophobic and hydrogen bonding networks. Protein dynamics shows that the overall structure, including loops, is very rigid and stabilized. By structural database analysis, we found that vEP C-ter100 shares its topology with that of the collagen-binding domain of collagenase, despite low sequence homology between the two domains. Fluorescence assay reveals that vEP C-ter100 interacts strongly with iron (Fe 3+ ). These findings suggest that vEP protease might recruit substrate molecules, such as collagen, by binding at C-ter100 and that vEP participates in iron uptake from iron-withholding proteins of the host cell during infection.",
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Solution structure and dynamics of C-terminal regulatory domain of Vibrio vulnificus extracellular metalloprotease. / Yun, Ji Hye; Kim, Heeyoun; Park, Jung Eun; Lee, Jung Sup; Lee, Weon Tae.

In: Biochemical and Biophysical Research Communications, Vol. 430, No. 2, 11.01.2013, p. 541-546.

Research output: Contribution to journalArticle

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