Insecticidal pilin subunit from the insect pathogen Xenorhabdus nematophila

Journal of Bacteriology
Puneet KhandelwalNirupama Banerjee

Abstract

Xenorhabdus nematophila is an insect pathogen and produces protein toxins which kill the larval host. Previously, we characterized an orally toxic, large, outer membrane-associated protein complex from the culture medium of X. nematophila. Here, we describe the cloning, expression, and characterization of a 17-kDa pilin subunit of X. nematophila isolated from that protein complex. The gene was amplified by PCR, cloned, and expressed in Escherichia coli. The recombinant protein was refolded in vitro in the absence of its cognate chaperone by using a urea gradient. The protein oligomerized during in vitro refolding, forming multimers. Point mutations in the conserved N-terminal residues of the pilin protein greatly destabilized its oligomeric organization, demonstrating the importance of the N terminus in refolding and oligomerization of the pilin subunit by donor strand complementation. The recombinant protein was cytotoxic to cultured Helicoverpa armigera larval hemocytes, causing agglutination and subsequent release of the cytoplasmic enzyme lactate dehydrogenase. The agglutination of larval cells by the 17-kDa protein was inhibited by several sugar derivatives. The biological activity of the purified recombinant protein indic...Continue Reading

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Citations

Jul 10, 2007·Nature Reviews. Microbiology·Erin E Herbert, Heidi Goodrich-Blair
Aug 1, 2008·The Journal of Biological Chemistry·Mohan Chandra JoshiNirupama Banerjee
Aug 23, 2011·Applied and Environmental Microbiology·Holly SnyderSteven Forst
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May 19, 2020·International Journal of Biological Macromolecules·Saquib MahmoodNeera Bhalla Sarin

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