Amylosin from Bacillus amyloliquefaeiens, a K+ and Na+ channel-forming toxic peptide containing a polyene structure

Raimo Mikkola*, Maria A. Andersson, Vera Teplova, Pavel Grigoriev, Till Kuehn, Sandra Loss, Irina Tsitko, Camelia Apetroaie, Nils-Erik L. Saris, Pirjo Veijalainen, Mirja S. Salkinoja-Salonen

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Bacillus amyloliquefaciens strains isolated from the indoor environment of moisture-damaged buildings produce a 1197 Da toxin, named amylosin. Nuclear magnetic resonance (NMR) data showed that amylosin contains a chromophoric polyene structure and the amino acids leucine/isoleucine, proline, aspartic acid/asparagine, glutamic acid/glutamine and tyrosine. A quantitation method for amylosin was developed using commercially available amphotericin B as a reference compound and a known concentration of amylosin determined by NMR with the electronic reference to access in vivo concentration (ERETIC) method. Purified amylosin inhibited motility of boar sperm cells at an exposure concentration of 135nM and hyperpolarized their cell membrane and depolarized their mitochondria at exposure to concentration of 33-67 nM for 10 min. In a 3-d exposure time only 27 nM of amylosin was needed to provoke the same toxicity functions. Amylosin was cytotoxic to feline lung cells at concentrations of <170 nM. Purified amylosin provoked adenosine 5'-triphosphate (ATP)-independent cation influx into isolated rat liver mitochondria (RLM), inducing swelling of the mitochondria, at concentrations of 200nm K+ or > 250nM Na+ medium. In the K+- or Na+-containing medium, amylosin uncoupled RLM, causing oxidation of pyridine nucleotides (PN), loss of the mitochondrial membrane potential, and suppressed ATP synthesis. Purified amylosin produced cation channels in black-lipid membranes (BLMs) with a selectivity K+ > Na+ at a concentration of 26 nM, i.e. the same concentration at which amylosin was toxic to boar sperm cells. The amylosin cation channels were cholesterol- and ATP-independent and more effective with K+ than with Na+.

Original languageEnglish
Pages (from-to)1158-1171
Number of pages14
JournalTOXICON
Volume49
Issue number8
DOIs
Publication statusPublished - 15 Jun 2007
MoE publication typeA1 Journal article-refereed

Keywords

  • amylosin
  • Bacillus amyloliquefaciens
  • channel
  • mitochondria
  • indoor air
  • swelling
  • ionophore
  • boar sperm
  • feline lung
  • uncoupling
  • RAT-LIVER MITOCHONDRIA
  • MACROLIDE ANTIBIOTICS
  • NMR STRUCTURE
  • SURFACTIN
  • MEMBRANE

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