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Home / Equine Research Bank / Infect Immun / Identification of eNAP-1, an antimicrobial peptide from equi...
Infection and immunity1992; 60(8); 3065-3071; doi: 10.1128/iai.60.8.3065-3071.1992

Identification of eNAP-1, an antimicrobial peptide from equine neutrophils.

Abstract: Endogenous, cysteine-rich antimicrobial peptides known as defensins are prominent components of human, rabbit, and rat neutrophils, yet little is known about their occurrence in other mammalian species. Although we did not detect mature (i.e., processed) defensins in equine neutrophil granules, we found that these granules contained small amounts of other cysteine-rich peptides with antimicrobial activity. One of these, eNAP-1, was purified by a combination of gel permeation and reversed-phase high-performance liquid chromatography from acid extracts prepared from the cytoplasmic granules of equine neutrophils. The molecular mass of eNAP-1 was approximately 7.2 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Amino acid analysis revealed that eNAP-1 had an unusually high cysteine content and that it was relatively enriched in alanine, glycine, lysine, and proline residues. The partial (N-terminal) amino acid sequence of eNAP-1 was DVQCGEGHFCHDXQTCCRASQGGXACCPYSQGVCCADQRHCCPVGF. Thirty-six of these residues (78.3%) were identical to those of a recently cloned human neutrophil peptide of unknown function and belonging to the granulin family. Homologous peptides have also been noted in rat bone marrow cells and rat kidney epithelins. We tested the ability of eNAP-1 to kill several equine uterine pathogens. Streptococcus zooepidemicus was killed most effectively, sustaining a greater than 99.8% decrease in CFU per milliliter after a 2-h exposure to 100 micrograms of eNAP-1 per ml (approximately 15 microM). Escherichia coli and Pseudomonas aeruginosa were somewhat less susceptible, manifesting 87.0 and 87.1% mean decreases in CFU per milliliter, respectively, after incubation for 2 h with 200 micrograms of eNAP-1 per ml. Klebsiella pneumoniae numbers were not significantly reduced after exposure to eNAP-1. These antimicrobial properties suggest that eNAP-1 may contribute to phagocyte-mediated host defense against equine infections.
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Publication Date: 1992-08-01 PubMed ID: 1639474PubMed Central: PMC257282DOI: 10.1128/iai.60.8.3065-3071.1992Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research focuses on the discovery of a new antimicrobial peptide, named eNAP-1, extracted from the neutrophil granules of horses. The study demonstrates eNAP-1’s ability to effectively combat certain equine pathogens.

Introduction and Methods

  • The study explores the occurrence of cysteine-rich antimicrobial peptides, known as defensins, in equine neutrophils. While such peptides have been identified in human, rabbit, and rat neutrophils, they have not been well-studied in other mammals.
  • Though mature defensins were not discovered in equine neutrophil granules, the researchers found similar peptides, including eNAP-1. This peptide was isolated and purified for this study.
  • To extract eNAP-1, the researchers combined gel permeation and reversed-phase high-performance liquid chromatography.

eNAP-1 Characteristics

  • An analysis showed that eNAP-1 had an unusually high cysteine content and was relatively enriched in alanine, glycine, lysine, and proline residues.
  • Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular mass of eNAP-1 was found to be approximately 7.2 kilodaltons.
  • In comparing the amino acid sequence of eNAP-1, researchers found it shared about 78.3% of the residues with a recently isolated human neutrophil peptide of the granulin family.

Antimicrobial Activity of eNAP-1

  • The researchers tested eNAP-1 against a variety of equine uterine pathogens. Streptococcus zooepidemicus was reduced by over 99.8% after a two-hour exposure to eNAP-1.
  • Escherichia coli and Pseudomonas aeruginosa showed somewhat lesser susceptibility, with respective reduction rates of 87.0% and 87.1% after two hours of exposure to eNAP-1.
  • Klebsiella pneumoniae numbers, however, did not significantly decrease after exposure to eNAP-1.

Conclusion

  • Scientists conclude that eNAP-1 has a key role in boosting phagocyte-aided defenses against specific infections in horses, yet further research is required to understand its total functions.

Cite This Article

APA
Couto MA, Harwig SS, Cullor JS, Hughes JP, Lehrer RI. (1992). Identification of eNAP-1, an antimicrobial peptide from equine neutrophils. Infect Immun, 60(8), 3065-3071. https://doi.org/10.1128/iai.60.8.3065-3071.1992

Publication

Infection and immunity
ISSN: 0019-9567
NlmUniqueID: 0246127
Country: United States
Language: English
Volume: 60
Issue: 8
Pages: 3065-3071

Researcher Affiliations

Couto, M A
  • Department of Reproduction, School of Veterinary Medicine, University of California, Davis 95616.
Harwig, S S
    Cullor, J S
      Hughes, J P
        Lehrer, R I

          MeSH Terms

          • Amino Acid Sequence
          • Amino Acids / analysis
          • Animals
          • Anti-Infective Agents / isolation & purification
          • Anti-Infective Agents / pharmacology
          • Base Sequence
          • Blood Proteins / isolation & purification
          • Blood Proteins / pharmacology
          • Defensins
          • Horses / blood
          • Molecular Sequence Data
          • Neutrophils / chemistry
          • Peptides / isolation & purification
          • Peptides / pharmacology

          Grant Funding

          • 1 S10RR05554-01 / NCRR NIH HHS
          • AI 22839 / NIAID NIH HHS

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