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Antimicrobial Agents and Chemotherapy, February 2003, p. 643-652, Vol. 47, No. 2
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.2.643-652.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

MUC7 20-Mer: Investigation of Antimicrobial Activity, Secondary Structure, and Possible Mechanism of Antifungal Action

Libuse A. Bobek* and Hongsa Situ

Department of Oral Biology, University at Buffalo, The State University of New York, Buffalo, New York 14214

Received 20 June 2002/ Returned for modification 4 September 2002/ Accepted 29 October 2002

This study was aimed at examining the spectrum of antimicrobial activity of MUC7 20-mer (N-LAHQKPFIRKSYKCLHKRCR-C; residues 32 to 51 of MUC7, the low-molecular-weight human salivary mucin, comprised of 357 residues) and comparing its antifungal properties to those of salivary histatin 5 (Hsn-5). We also examined the secondary structure of the 20-mer and the possible mechanism of its antifungal action. Our results showed that MUC7 20-mer displays potent killing activity against a variety of fungi and both gram-positive and gram-negative bacteria at micromolar concentrations. Time-dependent killing of Candida albicans and Cryptococcus neoformans by MUC7 20-mer and Hsn-5 indicated differences in killing rates between MUC7 20-mer and Hsn-5. The secondary structure prediction showed that MUC7 20-mer adopts an amphiphilic helix with distinguishable hydrophilic and hydrophobic faces (a characteristic that is associated with antimicrobial activity). In comparison to that of Hsn-5, the fungicidal activity of MUC7 20-mer against C. albicans seems to be independent of fungal cellular metabolic activity, as evidenced by its killing potency at a low temperature (4°C) and in the presence of inhibitors of oxidative phosphorylation in the mitochondrial system. Fluorescence microscopy showed the ability of MUC7 20-mer to cross the fungal cell membrane and to accumulate inside the cells. The internalization of MUC7 20-mer was inhibited by divalent cations. Confocal microscopy of cells doubly labeled with MUC7 20-mer and a mitochondrion-specific dye indicated that mitochondria are not the target of MUC7 20-mer for either C. albicans or C. neoformans.


* Corresponding author. Mailing address: Department of Oral Biology, State University of New York at Buffalo, 109 Foster Hall, 3435 Main St., Buffalo, NY 14214-3092. Phone: (716) 829-2465. Fax: (716) 829-3942. E-mail: lbobek{at}acsu.buffalo.edu.


Antimicrobial Agents and Chemotherapy, February 2003, p. 643-652, Vol. 47, No. 2
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.2.643-652.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.




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