A one-enzyme strategy to release an antimicrobial peptide from the LFampin-domain of bovine lactoferrin.
Antimicrobial peptides have been found throughout living nature, yet antimicrobial sequences may still lie hidden within a wide variety of proteins. A rational strategy was developed to select interesting domains, based on the presumed common features of antimicrobial peptides, and to release these from accessible and safe proteins. In silico proteolysis simulations of bovine lactoferrin (bLF) with selected endoproteinases predicted the liberation of peptides that encompasses a cationic amphipathic alpha-helix. Three predicted peptides were synthesized and tested for their biological activity, demonstrating that one single enzyme was sufficient to obtain an antimicrobial peptide. The proof of principle demonstrated that a 32-mer fragment isolated from the endoproteinase AspN digestion of bLF possessed strong antimicrobial activity. Moreover, desalted crude digest had improved activity over native bLF. Hence, selective digestion of bLF increases its antimicrobial activity by release of antimicrobial stretches.
Bolscher
Jan G M
JG
Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), Vrije Universiteit and Universiteit van Amsterdam, Van der Boechorststraat 7, NL-1081BT Amsterdam, The Netherlands. jgm.bolscher@vumc.nl
van der Kraan
Marieke I A
MI
Nazmi
Kamran
K
Kalay
Hakan
H
Grün
Christian H
CH
Van't Hof
Wim
W
Veerman
Enno C I
EC
Nieuw Amerongen
Arie V
AV
D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry / Q000378:metabolism*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000378:metabolism*; D002176:Candida albicans; D002417:Cattle; D019295:Computational Biology; D004926:Escherichia coli; D007781:Lactoferrin / Q000737:chemistry / Q000378:metabolism*; D007782:Lactoglobulins / Q000737:chemistry / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000737:chemistry / Q000502:physiology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary
D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D007782:Lactoglobulins; D010446:Peptide Fragments; C502040:lactoferrampin; D007781:Lactoferrin; D008666:Metalloendopeptidases; C058921:endoproteinase Asp-N