Ac-AP-12, a novel factor Xa anticoagulant peptide from the esophageal glands of adult Ancylostoma caninum

• Published in: Molecular and biochemical parasitology 2011-05, Vol.177 (1), p.42-48
• Main Authors: Jiang, Desheng, Zhan, Bin, Mayor, Reina S., Gillespie, Portia, Keegan, Brian, Bottazzi, Maria Elena, Hotez, Peter
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Ancylostoma - chemistry; Ancylostoma - classification; Ancylostoma - genetics; Ancylostoma - metabolism; Ancylostoma caninum; Ancylostomiasis - blood; Ancylostomiasis - parasitology; Animals; Anticoagulant; anticoagulants; Anticoagulants - chemistry; Anticoagulants - metabolism; Anticoagulants - pharmacology; Blood Coagulation - drug effects; Cloning, Molecular; Esophagus - chemistry; Esophagus - metabolism; Factor Xa Inhibitors; fXa; Humans; Molecular Sequence Data; Peptides - chemistry; Peptides - genetics; Peptides - metabolism; Peptides - pharmacology; Phylogeny; Sequence Alignment; Vaccine
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/j.molbiopara.2011.01.008

Ac-AP-12, an anticoagulant peptide with 9.1kDa produced by Ancylostoma caninum esophageal glands (es) exhibits potent anticoagulant activity (aPPT, PT) by inhibiting human fXa. [Display omitted] ► A novel anticoagulant peptide (Ac-AP-12) was cloned from Ancylostoma caninum. ► The recombinant protein demonstrated potent anticoagulant activity on human blood. ► It specifically inhibited human fXa activity. ► Immunolocalization showed its location at esophageal glands of adult hookworm. ► It may play an important role in the blood feeding and digestion. Immunoscreening an Ancylostoma caninum cDNA library with canine hookworm-infected dog serum resulted in the isolation of a 461bp cDNA encoding Ac-AP-12, a new 9.1kDa anticoagulant peptide (100 amino acids) with 43–69% amino acid homology to other nematode anticoagulant peptides (NAPs) from Ancylostoma hookworms. Messenger RNA transcription and expression of Ac-AP-12 was unique to the adult stage of A. caninum. The yeast expressed recombinant Ac-AP-12 demonstrated potent anticoagulant activity on human blood plasma in a concentration dependent manner, and was shown to specifically inhibit human factor Xa activity. Immunolocalization with specific rabbit antiserum showed that Ac-AP-12 was exclusively located in the esophageal glands of adult hookworm. Ac-AP-12 is hypothesized to facilitate both parasite blood feeding and digestion.