Human salivary MUC7 mucin peptides: effect of size, charge and cysteine residues on antifungal activity.
We have previously shown that MUC7 (human salivary low-molecular-mass mucin) 20-mer: LAHQKPFIRKSYKCLHKRCR (residues 32-51 of the parent MUC7, with a net positive charge of 7) possesses a broad-spectrum antimicrobial activity [Bobek and Situ (2003) Antimicrob. Agents Chemother. 47, 645-652]. The aims of the present study were to determine the minimum peptide chain length and its location within the 20-mer region that retains potent antifungal activity against Candida albicans and Cryptococcus neoformans and to examine the effect of net charge of the peptide as well as the role of cysteine residues on the fungicidal activity. First, several C-terminal truncated MUC7 20-mer fragments (16-mer, 12-mer, 11-mer, 10-mer and 8-mer) and one N-terminal fragment (8-mer-N) were synthesized and tested. The results showed that MUC7 12-mer, located at the C-terminal region of MUC7 20-mer, having a net charge of +6 and exhibiting an amphipathic helical conformation, not only retained but exceeded the antifungal activity of that of 20-mer. Secondly, several variants of the 12-mer peptide containing a lower or no net positive charge, or no cysteine residues were synthesized and tested. A clear correlation between the net positive charge of the 12-mer, its potency and initial interaction of peptide with fungal cells was found by killing assays, fluorescence microscopy and fungal cell-membrane potential measurements. Furthermore, cysteine residues, which play a critical role in bacterial binding, were found to be not important for the fungicidal activity of these peptides. These results identified MUC7 12-mer as a potential candidate for development into a novel antifungal therapeutic agent.
Situ
Hongsa
H
Department of Oral Biology, University at Buffalo, The State University of New York at Buffalo, 109 Foster Hall, 3435 Main Street, Buffalo, NY 14214-3092, USA.
Wei
Guoxian
G
Smith
Christina J
CJ
Mashhoon
Shirin
S
Bobek
Libuse A
LA
D000595:Amino Acid Sequence; D024361:Amino Acids, Basic / Q000737:chemistry; D000935:Antifungal Agents / Q000737:chemistry* / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects / Q000502:physiology; D002942:Circular Dichroism; D003455:Cryptococcus neoformans / Q000187:drug effects / Q000502:physiology; D003545:Cysteine / Q000737:chemistry / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry* / Q000494:pharmacology*; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D017433:Protein Structure, Secondary; D012471:Salivary Proteins and Peptides / Q000737:chemistry* / Q000494:pharmacology*; D055672:Static Electricity
D024361:Amino Acids, Basic; D000935:Antifungal Agents; C083238:MUC7 protein, human; D009077:Mucins; D010455:Peptides; D012471:Salivary Proteins and Peptides; D003545:Cysteine