Abstract
An anti-thrombin peptide (anophelin) was isolated from the salivary glands of the mosquito Anopheles albimanus through molecular sieving and reverse-phase high-performance liquid chromatography. The purified peptide inhibited thrombin-induced platelet aggregation, thrombin esterolytic activity on a synthetic substrate, and thrombin cleavage of fibrinogen. The purified anti-thrombin had a molecular mass of 6342.4 Da. Its amino terminus was blocked, but internal sequence yielded three peptide sequences, which were used to design oligonucleotide probes for polymerase chain reaction amplification of salivary gland cDNA and isolation of the full-length clone. Analysis of the sequence of anophelin shows no similarities to any other anti-thrombin peptides. Anophelin was successfully synthesized and characterized to be a tight-binding, specific, and novel inhibitor of thrombin.
MeSH terms
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Amino Acid Sequence
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Animals
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Anopheles / chemistry*
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Base Sequence
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Cloning, Molecular
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DNA, Complementary / isolation & purification
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Female
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Humans
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Insect Proteins / chemical synthesis
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Insect Proteins / genetics*
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Insect Proteins / isolation & purification*
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Insect Proteins / physiology
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Molecular Sequence Data
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Platelet Aggregation / drug effects
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Platelet Aggregation Inhibitors / chemical synthesis
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Platelet Aggregation Inhibitors / isolation & purification
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Platelet Aggregation Inhibitors / pharmacology
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Salivary Glands / chemistry
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Salivary Proteins and Peptides / chemical synthesis
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Salivary Proteins and Peptides / genetics*
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Salivary Proteins and Peptides / isolation & purification*
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Salivary Proteins and Peptides / physiology
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Thrombin / antagonists & inhibitors*
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Thrombin / pharmacology
Substances
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DNA, Complementary
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Insect Proteins
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Platelet Aggregation Inhibitors
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Salivary Proteins and Peptides
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anophelin protein, Anopheles albimanus
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Thrombin