Abstract
A carboxypeptidase inhibitor called HlTCI was isolated from Haemaphysalis longicornis in this study. The full-length cDNA of HlTCI contains an open reading frame (ORF) of 291 bp, encoding 96 amino acid residues consisting of a predicted 19-residue signal peptide and a putative mature 77-residue protein. The expected mature protein is cysteine-rich and has 12 cysteine residues assumed to construct six disulfide bridges. The deduced peptide sequence shows 63.9% homology to the carboxypeptidase inhibitor from another ixodid tick, Rhipicephalus bursa. Reverse-transcription PCR (RT-PCR) indicated that HlTCI was specifically expressed in the ovary from partially engorged adult ticks. The recombinant protein of HlTCI (rHlTCI) with glutathione S-transferase (GST) was expressed in Escherichia coli strain BL21 (DE3) and purified by glutathione-Sepharose 4B beads. rHlTCI showed inhibitory activity against digestive metallocarboxypeptidases A and B, but the activity was affected by the increase of the temperature treatment. High concentrations of rHlTCI were shown to significantly accelerate fibrinolysis in vitro. This effect of rHlTCI on clot lysis suggests its promising potential for use in some thrombotic disorders.
| Original language | English |
|---|---|
| Pages (from-to) | 1079-1087 |
| Number of pages | 9 |
| Journal | Journal of Insect Physiology |
| Volume | 53 |
| Issue number | 10 |
| DOIs | |
| Publication status | Published - Oct 1 2007 |
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All Science Journal Classification (ASJC) codes
- Insect Science
- Physiology
Cite this
Characterization of a carboxypeptidase inhibitor from the tick Haemaphysalis longicornis. / Gong, Haiyan; Zhou, Jinlin; Liao, Min; Hatta, Takeshi; Harnnoi, Thasaneeya; Umemiya, Rika; Inoue, Noboru; Xuan, Xuenan; Fujisaki, Kozo.
In: Journal of Insect Physiology, Vol. 53, No. 10, 01.10.2007, p. 1079-1087.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of a carboxypeptidase inhibitor from the tick Haemaphysalis longicornis
AU - Gong, Haiyan
AU - Zhou, Jinlin
AU - Liao, Min
AU - Hatta, Takeshi
AU - Harnnoi, Thasaneeya
AU - Umemiya, Rika
AU - Inoue, Noboru
AU - Xuan, Xuenan
AU - Fujisaki, Kozo
PY - 2007/10/1
Y1 - 2007/10/1
N2 - A carboxypeptidase inhibitor called HlTCI was isolated from Haemaphysalis longicornis in this study. The full-length cDNA of HlTCI contains an open reading frame (ORF) of 291 bp, encoding 96 amino acid residues consisting of a predicted 19-residue signal peptide and a putative mature 77-residue protein. The expected mature protein is cysteine-rich and has 12 cysteine residues assumed to construct six disulfide bridges. The deduced peptide sequence shows 63.9% homology to the carboxypeptidase inhibitor from another ixodid tick, Rhipicephalus bursa. Reverse-transcription PCR (RT-PCR) indicated that HlTCI was specifically expressed in the ovary from partially engorged adult ticks. The recombinant protein of HlTCI (rHlTCI) with glutathione S-transferase (GST) was expressed in Escherichia coli strain BL21 (DE3) and purified by glutathione-Sepharose 4B beads. rHlTCI showed inhibitory activity against digestive metallocarboxypeptidases A and B, but the activity was affected by the increase of the temperature treatment. High concentrations of rHlTCI were shown to significantly accelerate fibrinolysis in vitro. This effect of rHlTCI on clot lysis suggests its promising potential for use in some thrombotic disorders.
AB - A carboxypeptidase inhibitor called HlTCI was isolated from Haemaphysalis longicornis in this study. The full-length cDNA of HlTCI contains an open reading frame (ORF) of 291 bp, encoding 96 amino acid residues consisting of a predicted 19-residue signal peptide and a putative mature 77-residue protein. The expected mature protein is cysteine-rich and has 12 cysteine residues assumed to construct six disulfide bridges. The deduced peptide sequence shows 63.9% homology to the carboxypeptidase inhibitor from another ixodid tick, Rhipicephalus bursa. Reverse-transcription PCR (RT-PCR) indicated that HlTCI was specifically expressed in the ovary from partially engorged adult ticks. The recombinant protein of HlTCI (rHlTCI) with glutathione S-transferase (GST) was expressed in Escherichia coli strain BL21 (DE3) and purified by glutathione-Sepharose 4B beads. rHlTCI showed inhibitory activity against digestive metallocarboxypeptidases A and B, but the activity was affected by the increase of the temperature treatment. High concentrations of rHlTCI were shown to significantly accelerate fibrinolysis in vitro. This effect of rHlTCI on clot lysis suggests its promising potential for use in some thrombotic disorders.
UR - http://www.scopus.com/inward/record.url?scp=34748904625&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34748904625&partnerID=8YFLogxK
U2 - 10.1016/j.jinsphys.2007.06.008
DO - 10.1016/j.jinsphys.2007.06.008
M3 - Article
C2 - 17651749
AN - SCOPUS:34748904625
VL - 53
SP - 1079
EP - 1087
JO - Journal of Insect Physiology
JF - Journal of Insect Physiology
SN - 0022-1910
IS - 10
ER -