The role of the abundant phenylalanines in the mode of action of the antimicrobial peptide clavanin.
Clavanin A is a special antimicrobial peptide that acts at the level of the membrane via a pH-dependent mechanism. At neutral pH, clavanin disrupts biological and model membranes in a nonspecific manner, causing efflux of large molecules. At mildly acidic conditions, however, the peptide efficiently kills bacteria by permeabilizing their membrane most likely by interacting with proteins involved in proton translocation [Biochemistry 41 (2002) 7529]. Clavanin A is unusually rich in phenylalanines with 5 out of 23 residues, which suggests that these residues are functionally important. A set of mutants, in which all Phe residues are replaced by either Ile, Leu, Trp, or Tyr was used to investigate the role of these amino acids. The antimicrobial activities of the different peptides both at neutral and low pH show that the presence of phenylalanine is not essential nor optimal, as the Trp, Leu, and Ile mutant are equally or more active than the wild-type component. In general, at neutral pH, the biological activities correlate well with the peptides' ability to interact with membrane lipids. Correspondingly, the permeabilization efficiencies of biological and model membranes of the various derivatives were found to be closely related to their ability to adopt alpha-helical structures, and follows the order 5L>5W>5I>5Y>wild type. The results suggest an important role for the Phe residues, in providing the peptide in a balanced manner with sufficient hydrophobicity, and therewith membrane affinity, as well as conformational flexibility.
van Kan
Ellen J M
EJ
Department of Functional Ingredients, Food and Food Processing, Agrotechnological Research Institute (ATO B.V.), Wageningen University and Research Centre, Bornsesteeg 59, 6708 PD Wageningen, The Netherlands. ellen.vankan@wur.nl
Demel
Rudy A
RA
van der Bent
Arie
A
de Kruijff
Ben
B
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology; D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism*; D002942:Circular Dichroism; D008837:Micrococcus / Q000187:drug effects / Q000378:metabolism; D010649:Phenylalanine / Q000378:metabolism*; D011817:Rabbits
D000900:Anti-Bacterial Agents; D001798:Blood Proteins; C106713:clavanin A; D010649:Phenylalanine