eNAP-2, a novel cysteine-rich bactericidal peptide from equine leukocytes

M. A. Couto, S. S L Harwig, James S Cullor, J. P. Hughes, R. I. Lehrer

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Abstract

We purified a novel cysteine-rich antibiotic peptide, eNAP-2 (M(r), ~6,500), from acid extracts of equine neutrophils by sequential gel filtration and reversed-phase high-performance liquid chromatography and determined its partial N-terminal amino acid sequence. Although its cysteine motif distinguished eNAP-2 from all other currently known endogenous antibiotic peptides, including defensins and granulins, it showed substantial sequence similarity to WDNM1, a putative member of the four-disulfide-core protein family that also includes animal and human antiproteases, snake venom neurotoxins, and rat and mouse whey proteins. The antibacterial properties of eNAP-2 were tested against several equine uterine pathogens, namely, Streptococcus zooepidemicus, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae. Killing of S. zooepidemicus was very efficient, as evidenced by a 94% decrease in numbers of CFU per milliliter after exposure to 100 μg of eNAP-2 per ml (~15 μM) for 2 h. Exposure of E. coli and P. aeruginosa to 200 μg of eNAP-2 per ml for 2 h resulted in 90.2 and 77.6% reduction, respectively, in the numbers of CFU per milliliter. Bacteriostasis, without bactericidal activity, occurred after K. pneumoniae was incubated with 200 μg of eNAP-2 per ml. Additional studies will be required in other species and cell types to determine whether eNAP-2 is restricted to equine neutrophils or is the index member of a larger family of endogenous antibiotics.

Original languageEnglish (US)
Pages (from-to)5042-5047
Number of pages6
JournalInfection and Immunity
Volume60
Issue number12
StatePublished - 1992

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ASJC Scopus subject areas

  • Immunology

Cite this

Couto, M. A., Harwig, S. S. L., Cullor, J. S., Hughes, J. P., & Lehrer, R. I. (1992). eNAP-2, a novel cysteine-rich bactericidal peptide from equine leukocytes. Infection and Immunity, 60(12), 5042-5047.