Antimicrobial activity of a chelatable poly(arginyl-histidine) produced by the ergot fungus Verticillium kibiense.
We have recently developed a convenient method of screening a broad range of microorganisms that produce epsilon-poly-L-lysine (M. Nishikawa and K. Ogawa, Appl. Environ. Microbiol. 68:3575-3581, 2002). Using this method, we found an ergot fungus that secretes a charged polypeptide other than epsilon-poly-L-lysine. It was identified as a new species on the basis of its 28S rRNA sequence and was named Verticillium kibiense (formerly Epichloe kibiensis). Peptide sequencing and mass spectrometry revealed that the polypeptide is a linear peptide composed of repeated units of arginyl-histidine. The numbers of repeated units were in most cases five and in some cases four or six. This peptide showed activity against a broad range of bacteria and fungi but lost its activity under conditions of high ionic strength. Zinc and copper ions specifically changed the circular dichroism spectra of the peptide and restored the antimicrobial activity from abrogation under high ionic conditions, although these ions had no reinforcing effect on antimicrobial activity when they were added to solutions at a low ionic strength. The peptide labeled with fluorescein was able to permeate the cell membranes of target microbes, but its ability to permeate cell membranes decreased under conditions of high ionic strength. This decreased ability was partially recovered specifically by the addition of zinc and copper ions. These results indicate that poly(arginyl-histidine) is a cationic polypeptide characterized by specific metal binding and resistance to salts.
Nishikawa
Masanobu
M
Research Institute for Biological Sciences, Okayama (RIBS), 7549-1 Kayo-cho, Okayama 716-1241, Japan. mnishikawa@bio-ribs.com
Ogawa
Ken'ichi
K
D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects*; D002462:Cell Membrane / Q000187:drug effects; D002614:Chelating Agents / Q000737:chemistry*; D002942:Circular Dichroism; D004912:Erythrocytes / Q000187:drug effects; D006461:Hemolysis / Q000187:drug effects; D066298:In Vitro Techniques; D008081:Liposomes / Q000737:chemistry; D008670:Metals / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008856:Microscopy, Fluorescence; D010455:Peptides / Q000494:pharmacology; D012756:Sheep; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D020172:Verticillium / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D002614:Chelating Agents; D008081:Liposomes; D008670:Metals; D010455:Peptides