title: Interactions of monomeric rabbit neutrophil defensins with bilayers: comparison with dimeric human defensin HNP-2.
abstract: Human antimicrobial neutrophil defensin HNP-2 has been shown to form large multimeric pores in pure 1-palmitoyl-2-oleoyl phosphatidylglycerol (POPG) bilayers that lead to all-or-none release of vesicle contents [Wimley et al. (1994) Protein Sci.3, 1362-1373]. Because human neutrophil defensins form natural dimers in solution, the question arises as to the role of dimerization in pore formation. However, the dimers are so stable that this question is not easily answered directly. Rabbit neutrophil defensins, whose three-dimensional structures are very similar to those of human defensins, are monomeric in aqueous solution and thus provide an opportunity to test the hypothesis that dimerization may play a role in multimeric pore formation. We therefore examined the interactions of the six known rabbit neutrophil defensins with large unilamellar vesicles (LUV) under the conditions known to lead to stable pore formation by HNP-2. We find that the rabbit defensins bind strongly to LUVs formed from pure POPG or mixtures of POPG with neutral (zwitterionic) phospholipid but induce leakage of vesicle contents only from pure POPG vesicles. Rabbit defensin NP-4 does not cause leakage under any conditions examined. The remaining defensins, NP-1, NP-2, NP-3A, NP-3B, and NP-5, cause graded release of the contents of pure POPG vesicles as does a mixture of the six defensins. The graded release indicates that the rabbit defensins do not form stable pores in the membrane. This result thus suggests that the structural features of human defensins that permit dimer formation in aqueous solution are likely to be important in the formation of multimeric pores.
authors.affiliation: Department of Physiology and Biophysics, University of California, Irvine 92697-4560, USA.
authors.forename: K
authors.initials: K
authors.lastname: Hristova
authors.forename: M E
authors.initials: ME
authors.lastname: Selsted
authors.forename: S H
authors.initials: SH
authors.lastname: White
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects; D017124:Conserved Sequence; D023082:Defensins; D005453:Fluorescence; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009281:Naphthalenes / Q000378:metabolism; D009504:Neutrophils / Q000737:chemistry*; D010539:Permeability / Q000187:drug effects; D010713:Phosphatidylcholines / Q000032:analysis; D010715:Phosphatidylglycerols / Q000032:analysis; D011487:Protein Conformation; D011726:Pyridinium Compounds / Q000378:metabolism; D011817:Rabbits; D023084:alpha-Defensins
chemical_list: D001798:Blood Proteins; D023082:Defensins; D008051:Lipid Bilayers; D008081:Liposomes; D009281:Naphthalenes; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D011726:Pyridinium Compounds; D023084:alpha-Defensins; C406566:defensin NP-3a; C406416:human neutrophil peptide 2; C406567:peptide NP-3b; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; C046465:8-amino-1,3,6-naphthalenetrisulfonic acid