Plantaricin A, a cationic peptide produced by Lactobacillus plantarum, permeabilizes eukaryotic cell membranes by a mechanism dependent on negative surface charge linked to glycosylated membrane proteins

Biochim Biophys Acta. 2013 Feb;1828(2):249-59. doi: 10.1016/j.bbamem.2012.11.001. Epub 2012 Nov 8.

Abstract

Lactobacillus plantarum C11 releases plantaricin A (PlnA), a cationic peptide pheromone that has a membrane-permeabilizing, antimicrobial effect. We have previously shown that PlnA may also permeabilize eukaryotic cells, with a potency that differs between cell types. It is generally assumed that cationic antimicrobial peptides exert their effects through electrostatic attraction to negatively charged phospholipids in the membrane. The aim of the present study was to investigate if removal of the negative charge linked to glycosylated proteins at the cell surface reduces the permeabilizing potency of PlnA. The effects of PlnA were tested on clonal rat anterior pituitary cells (GH(4) cells) using patch clamp and microfluorometric techniques. In physiological extracellular solution, GH(4) cells are highly sensitive to PlnA, but the sensitivity was dramatically reduced in solutions that partly neutralize the negative surface charge of the cells, in agreement with the notion that electrostatic interactions are probably important for the PlnA effects. Trypsination of cells prior to PlnA exposure also rendered the cells less sensitive to the peptide, suggesting that negative charges linked to membrane proteins are involved in the permeabilizing action. Finally, pre-exposure of cells to a mixture of enzymes that split carbohydrate residues from the backbone of glycosylated proteins also impeded the PlnA-induced membrane permeabilization. We conclude that electrostatic attraction between PlnA and glycosylated membrane proteins is probably an essential first step before PlnA can interact with membrane phospholipids. Deviating glycosylation patterns may contribute to the variation in PlnA sensitivity of different cell types, including cancerous cells and their normal counterparts.

MeSH terms

  • Animals
  • Bacteriocins / chemistry*
  • Calcium / chemistry
  • Cell Membrane Permeability
  • Cytophotometry / methods
  • Electrophysiology / methods
  • Glycosylation
  • Lactobacillus plantarum / chemistry*
  • Membrane Proteins / chemistry
  • Peptides / chemistry*
  • Phospholipids / chemistry
  • Pituitary Gland, Anterior / metabolism
  • Rats
  • Static Electricity
  • Surface Properties
  • Trypsin / pharmacology

Substances

  • Bacteriocins
  • Membrane Proteins
  • Peptides
  • Phospholipids
  • plantaricin A
  • Trypsin
  • Calcium