Cloning of habutobin cDNA and antithrombotic activity of recombinant protein

The habutobin cDNA was cloned from total RNA extracted from venom glands of Trimeresurus flavoviridis (the habu snake). The conceptual translation of 1539 bp of habutobin cDNA consists of 236 amino acids and its molecular weight is 25.7 kDa. Histidine (His)-tagged recombinant habutobin fusion protei...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2007-11, Vol.362 (4), p.899-904
Main Authors: Sunagawa, Masanori, Nakamura, Mariko, Kosugi, Tadayoshi
Format: Article
Language:English
Subjects:
60 APPLIED LIFE SCIENCES
AGGLOMERATION
Amino Acid Sequence
Animals
cDNA cloning
CLONING
Cloning, Molecular - methods
COLLAGEN
DNA, Complementary - genetics
DRUGS
FIBRIN
Fibrin forming activity
Fibrinolytic Agents - administration & dosage
Fibrinolytic Agents - metabolism
GLANDS
Habutobin
HISTIDINE
Inhibition of platelet aggregation
Molecular Sequence Data
MOLECULAR WEIGHT
MONOMERS
Platelet Aggregation - drug effects
RABBITS
Recombinant protein
Recombinant Proteins - administration & dosage
Recombinant Proteins - metabolism
RNA
Serine Endopeptidases - administration & dosage
Serine Endopeptidases - genetics
Serine Endopeptidases - metabolism
SNAKES
STRUCTURE FUNCTIONS
THROMBIN
Thrombin-like enzyme
Trimeresurus flavoviridis
VENOMS
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Summary:The habutobin cDNA was cloned from total RNA extracted from venom glands of Trimeresurus flavoviridis (the habu snake). The conceptual translation of 1539 bp of habutobin cDNA consists of 236 amino acids and its molecular weight is 25.7 kDa. Histidine (His)-tagged recombinant habutobin fusion protein, pET-r-habutobin and AcNPV-r-habutobin, was purified by bacterial system and baculoviral system, respectively. After refolding pET-r-habutobin, there were two protein bands at about 32 kDa and 65 kDa, indicating that habutobin might be produced as a monomer protein and processed to form two concatenated protein. Purified AcNPV-r-habutobin dose-dependently increased fibrin forming activity and inhibited collagen-induced aggregation of rabbit washed platelets. Thus, AcNPV-r-habutobin produced by baculoviral system is very useful for study on structure–function relationship, which is necessary for developing an antithrombotic drug from habutobin.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2007.08.103