Selective reciprocity in antimicrobial activity versus cytotoxicity of hBD-2 and crotamine

Edited by H. Ronald Kaback, University of California, Los Angeles, CA, and approved June 24, 2009
September 1, 2009
106 (35) 14972-14977

Abstract

Recent discoveries suggest cysteine-stabilized toxins and antimicrobial peptides have structure–activity parallels derived by common ancestry. Here, human antimicrobial peptide hBD-2 and rattlesnake venom-toxin crotamine were compared in phylogeny, 3D structure, target cell specificity, and mechanisms of action. Results indicate a striking degree of structural and phylogenetic congruence. Importantly, these polypeptides also exhibited functional reciprocity: (i) they exerted highly similar antimicrobial pH optima and spectra; (ii) both altered membrane potential consistent with ion channel-perturbing activities; and (iii) both peptides induced phosphatidylserine accessibility in eukaryotic cells. However, the Nav channel-inhibitor tetrodotoxin antagonized hBD-2 mechanisms, but not those of crotamine. As crotamine targets eukaryotic ion channels, computational docking was used to compare hBD-2 versus crotamine interactions with prototypic bacterial, fungal, or mammalian Kv channels. Models support direct interactions of each peptide with Kv channels. However, while crotamine localized to occlude Kv channels in eukaryotic but not prokaryotic cells, hBD-2 interacted with prokaryotic and eukaryotic Kv channels but did not occlude either. Together, these results support the hypothesis that antimicrobial and cytotoxic polypeptides have ancestral structure-function homology, but evolved to preferentially target respective microbial versus mammalian ion channels via residue-specific interactions. These insights may accelerate development of anti-infective or therapeutic peptides that selectively target microbial or abnormal host cells.

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Acknowledgments.

We thank Trang Phan and Scott G. Filler (Division of Infectious Diseases, Harbor–UCLA Medical Center, and the General Clinical Research Center at Harbor–UCLA Medical Center) for providing HUVECs for these studies and H. Ronald Kaback, Terry J. Smith, Robert I. Lehrer, Eric P. Brass, and John E. Edwards, Jr., for helpful discussions. This work was supported by National Institutes of Health, National Institute of Allergy and Infectious Diseases Grants 5R01AI39001 and 5R01AI48031 to M.R.Y.

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Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 106 | No. 35
September 1, 2009
PubMed: 19706485

Classifications

Submission history

Received: April 23, 2009
Published online: September 1, 2009
Published in issue: September 1, 2009

Keywords

  1. channel
  2. defensin
  3. host defense
  4. toxin

Acknowledgments

We thank Trang Phan and Scott G. Filler (Division of Infectious Diseases, Harbor–UCLA Medical Center, and the General Clinical Research Center at Harbor–UCLA Medical Center) for providing HUVECs for these studies and H. Ronald Kaback, Terry J. Smith, Robert I. Lehrer, Eric P. Brass, and John E. Edwards, Jr., for helpful discussions. This work was supported by National Institutes of Health, National Institute of Allergy and Infectious Diseases Grants 5R01AI39001 and 5R01AI48031 to M.R.Y.

Notes

This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/cgi/content/full/0904465106/DCSupplemental.

Authors

Affiliations

Nannette Y. Yount
Division of Infectious Diseases, LAC-Harbor University of California, Los Angeles Medical Center, Torrance, CA 90509;
St. John's Cardiovascular Research Center, Los Angeles Biomedical Research Institute at Harbor–University of California, Los Angeles, Torrance, CA 90502;
Deborah Kupferwasser
Division of Infectious Diseases, LAC-Harbor University of California, Los Angeles Medical Center, Torrance, CA 90509;
St. John's Cardiovascular Research Center, Los Angeles Biomedical Research Institute at Harbor–University of California, Los Angeles, Torrance, CA 90502;
Alberto Spisni
Department of Experimental Medicine, Section of Chemistry and Structural Biology, University of Parma, 43100 Parma, Italy;
Stephen M. Dutz
Department of Chemistry and Center for Macromolecular Modeling and Materials Design, California State Polytechnic University, Pomona, CA 91768; and
Zachary H. Ramjan
Department of Chemistry and Center for Macromolecular Modeling and Materials Design, California State Polytechnic University, Pomona, CA 91768; and
Shantanu Sharma
Department of Chemistry and Center for Macromolecular Modeling and Materials Design, California State Polytechnic University, Pomona, CA 91768; and
Alan J. Waring
Department of Medicine, David Geffen School of Medicine at University of California, Los Angeles, CA 90024
Michael R. Yeaman1 [email protected]
Division of Infectious Diseases, LAC-Harbor University of California, Los Angeles Medical Center, Torrance, CA 90509;
St. John's Cardiovascular Research Center, Los Angeles Biomedical Research Institute at Harbor–University of California, Los Angeles, Torrance, CA 90502;
Department of Medicine, David Geffen School of Medicine at University of California, Los Angeles, CA 90024

Notes

1
To whom correspondence should be addressed. E-mail: [email protected]
Author contributions: N.Y.Y. and M.R.Y. designed research; N.Y.Y., D.K., S.M.D., Z.H.R., S.S., A.J.W., and M.R.Y. performed research; N.Y.Y., A.S., S.S., and M.R.Y. contributed new reagents/analytic tools; N.Y.Y., S.M.D., Z.H.R., S.S., A.J.W., and M.R.Y. analyzed data; and N.Y.Y., A.S., S.S., A.J.W., and M.R.Y. wrote the paper.

Competing Interests

Conflict of interest statement: M.R.Y. is a shareholder of NovaDigm Therapeutics, Inc., and has received research funding from Pfizer, Inc., Amgen, Inc., Cubist Pharmaceuticals, and Novozymes Pharmaceuticals. None of these entities provided support for the current studies. This article is a PNAS Direct Submission.

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    Selective reciprocity in antimicrobial activity versus cytotoxicity of hBD-2 and crotamine
    Proceedings of the National Academy of Sciences
    • Vol. 106
    • No. 35
    • pp. 14735-15091

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