Abstract
Vibrio extracellular metalloprotease (vEP), secreted from Vibrio vulnificus, shows various proteolytic function such as prothrombin activation and fibrinolytic activities. Premature form of vEP has an N-terminal (nPP) and a C-terminal (C-ter100) region. The nPP and C-ter100 regions are autocleaved for the matured metalloprotease activity. It has been proposed that two regions play a key role in regulating enzymatic activity of vEP. Especially, C-ter100 has a regulatory function on proteolytic activity of vEP. C-ter100 domain has been cloned into the E. coli expression vectors, pET 32a and pGEX 4T-1 with TEV protease cleavage site and purified using gel-filtration chromatography followed by affinity chromatography. To understand how C-ter100 modulates proteolytic activity of vEP, structural studies were performed by heteronuclar multi-dimensional NMR spectroscopy. Backbone 1H, 15N and 13C resonances were assigned by data from standard triple resonance and HCCH-TOCSY experiments. The secondary structures of vEP C-ter100 were determined by TALOS+ and CSI software based on hydrogen/deuterium exchange. NMR data show that C-ter100 of vEP forms a β-barrel structure consisting of eight β-strands.
| Original language | English |
|---|---|
| Pages (from-to) | 3248-3252 |
| Number of pages | 5 |
| Journal | Bulletin of the Korean Chemical Society |
| Volume | 33 |
| Issue number | 10 |
| DOIs | |
| Publication status | Published - 2012 Oct 20 |
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All Science Journal Classification (ASJC) codes
- Chemistry(all)
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1H, 15N and 13C backbone assignments and secondary structures of c-ter100 domain of vibrio extracellular metalloprotease derived from vibrio vulnificus. / Yun, Ji Hye; Kim, Heeyoun; Park, Jung Eun; Cheong, Hae Kap; Cheong, Chaejoon; Lee, Jung Sup; Lee, Weontae.
In: Bulletin of the Korean Chemical Society, Vol. 33, No. 10, 20.10.2012, p. 3248-3252.Research output: Contribution to journal › Article
TY - JOUR
T1 - 1H, 15N and 13C backbone assignments and secondary structures of c-ter100 domain of vibrio extracellular metalloprotease derived from vibrio vulnificus
AU - Yun, Ji Hye
AU - Kim, Heeyoun
AU - Park, Jung Eun
AU - Cheong, Hae Kap
AU - Cheong, Chaejoon
AU - Lee, Jung Sup
AU - Lee, Weontae
PY - 2012/10/20
Y1 - 2012/10/20
N2 - Vibrio extracellular metalloprotease (vEP), secreted from Vibrio vulnificus, shows various proteolytic function such as prothrombin activation and fibrinolytic activities. Premature form of vEP has an N-terminal (nPP) and a C-terminal (C-ter100) region. The nPP and C-ter100 regions are autocleaved for the matured metalloprotease activity. It has been proposed that two regions play a key role in regulating enzymatic activity of vEP. Especially, C-ter100 has a regulatory function on proteolytic activity of vEP. C-ter100 domain has been cloned into the E. coli expression vectors, pET 32a and pGEX 4T-1 with TEV protease cleavage site and purified using gel-filtration chromatography followed by affinity chromatography. To understand how C-ter100 modulates proteolytic activity of vEP, structural studies were performed by heteronuclar multi-dimensional NMR spectroscopy. Backbone 1H, 15N and 13C resonances were assigned by data from standard triple resonance and HCCH-TOCSY experiments. The secondary structures of vEP C-ter100 were determined by TALOS+ and CSI software based on hydrogen/deuterium exchange. NMR data show that C-ter100 of vEP forms a β-barrel structure consisting of eight β-strands.
AB - Vibrio extracellular metalloprotease (vEP), secreted from Vibrio vulnificus, shows various proteolytic function such as prothrombin activation and fibrinolytic activities. Premature form of vEP has an N-terminal (nPP) and a C-terminal (C-ter100) region. The nPP and C-ter100 regions are autocleaved for the matured metalloprotease activity. It has been proposed that two regions play a key role in regulating enzymatic activity of vEP. Especially, C-ter100 has a regulatory function on proteolytic activity of vEP. C-ter100 domain has been cloned into the E. coli expression vectors, pET 32a and pGEX 4T-1 with TEV protease cleavage site and purified using gel-filtration chromatography followed by affinity chromatography. To understand how C-ter100 modulates proteolytic activity of vEP, structural studies were performed by heteronuclar multi-dimensional NMR spectroscopy. Backbone 1H, 15N and 13C resonances were assigned by data from standard triple resonance and HCCH-TOCSY experiments. The secondary structures of vEP C-ter100 were determined by TALOS+ and CSI software based on hydrogen/deuterium exchange. NMR data show that C-ter100 of vEP forms a β-barrel structure consisting of eight β-strands.
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U2 - 10.5012/bkcs.2012.33.10.3248
DO - 10.5012/bkcs.2012.33.10.3248
M3 - Article
AN - SCOPUS:84867887577
VL - 33
SP - 3248
EP - 3252
JO - Bulletin of the Korean Chemical Society
JF - Bulletin of the Korean Chemical Society
SN - 0253-2964
IS - 10
ER -