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Hookworm NK cell binding protein: identifying the 80kDa protein in Necator americanus excretory/secretory products
Author(s) -
Gaze Soraya,
Constant Stephanie,
Bethony Jeff,
Smout Michael,
Loukas Alex
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.674.5
Subject(s) - necator americanus , biotinylation , biology , flow cytometry , biochemistry , immunology , microbiology and biotechnology , helminths , ascaris lumbricoides
Hookworms are blood‐feeding parasites which can infect mammals, including humans, and heavy infection causes iron deficiency anaemia and malnutrition. Although humans usually acquire resistance to parasitic worms, this is not clear cut in human hookworm infections. Some nematodes, including N. americanus , are able to produce excretory/secretory (E/S) proteins and these can be collected from live adult worms by culturing them for short periods in vitro. These E/S proteins have demonstrated different functions including suppression of cellular proliferation, proteolytic activity and mimicry of chemokines. Our group previously showed that E/S proteins from N. americanus can bind to NK cells, induce IFN‐γ release and is bigger than 50kDa. This protein was called NKBP (NK cell binding protein), and in this work, we have begun to identify this protein using anion exchange chromatography with different salt gradients. After biotinylation, the E/S proteins were separated using chromatography, and fractions were monitored for NKBP by flow cytometry. These positive fractions were pooled and submitted to anion exchange chromatography using different salt gradient, and further purified to a fraction containing two major biotinylated proteins. In this work we show a potential protein around 80kDa and we are now in the process of identifying these proteins using tandem mass spectrometry. Hookworm E/S proteins are critical in allowing these parasites to survive in a hostile microenvironment, and elucidation of the roles of these molecules will aid in the development of new cures for hookworm disease as well as novel therapeutics for human autoimmune disorders.