Phase I of the planned program consists of taking a summer course in molecular biology and auditing five courses. Phases I and II will include different portions of the research program. Human hookworm infection is characterized by chronic GI bleeding and iron deficiency anemia. The adult worm facilitates blood feeding through secretion of a potent anticoagulant called the Hookworm Extrinsic Pathway Inhibitor (HEPI). The candidate has already isolated this novel 16 kDa protein from lysates of Ancylostoma duodenale. Synthetic oligonucleotides from amino acid sequences generated from tryptic digests of the protein have been used to generate and clone a PCR product containing a partial coding region sequence. The candidate plans to clone the full length gene from a cDNA library, to sequence this and to express recombinant HEPI in a bacterial or mammalian vector system. This expressed protein will be used to generate antibody for immunolocallization of the protein in the organism and for biochemical assays of protein activity.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Physician Scientist Award (K11)
Project #
5K11AI001299-04
Application #
2671368
Study Section
Microbiology and Infectious Diseases B Subcommittee (MID)
Project Start
1995-09-15
Project End
2000-08-31
Budget Start
1998-09-01
Budget End
1999-08-31
Support Year
4
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Yale University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Milstone, A M; Harrison, L M; Bungiro, R D et al. (2000) A broad spectrum Kunitz type serine protease inhibitor secreted by the hookworm Ancylostoma ceylanicum. J Biol Chem 275:29391-9
Donnelly, K M; Bromberg, M E; Milstone, A et al. (1998) Ancylostoma caninum anticoagulant peptide blocks metastasis in vivo and inhibits factor Xa binding to melanoma cells in vitro. Thromb Haemost 79:1041-7
Cappello, M; Hawdon, J M; Jones, B F et al. (1996) Ancylostoma caninum anticoagulant peptide: cloning by PCR and expression of soluble, active protein in E. coli. Mol Biochem Parasitol 80:113-7