HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Edgerton, M. Articles by Raj, P. A. Search for Related Content PubMed PubMed Citation Articles by Edgerton, M. Articles by Raj, P. A. Social Bookmarking                      What's this?

J Biol Chem, Vol. 273, Issue 32, 20438-20447, August 7, 1998

Candidacidal Activity of Salivary Histatins
IDENTIFICATION OF A HISTATIN 5-BINDING PROTEIN ON Candida albicans

Mira Edgerton^§, Svetlana E. Koshlukova, Thomas E. Lo, Brian G. Chrzan, Robert M. Straubinger, and Periathamby A. Raj^**

From the Departments of  Oral Biology and ^§ Restorative Dentistry, School of Dental Medicine and the  Department of Pharmaceutics, State University of New York, Buffalo, New York 14214 and the ^** Division of Toxicology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

Candida albicans is the predominant species of yeast isolated from patients with oral candidiasis, which is frequently a symptom of human immunodeficiency virus infection and is a criterion for staging and progression of AIDS. Salivary histatins (Hsts) are potent in vitro antifungal agents and have great promise as therapeutic agents in humans with oral candidiasis. The molecular mechanisms by which Hsts kill yeast cells are not known. We report here, that unlike other antimicrobial proteins, Hsts do not display lytic activities to lipid membranes, measured by release and dequenching of the fluorescent dye calcein. Analysis of the magnitude and time course of Hst-induced calcein release from C. albicans cells further showed that loss of cell integrity was a secondary effect following cell death, rather than the result of primary disruption of the yeast cell membrane. ¹²⁵I-Hst 5 binding studies indicated that C. albicans expressed a class of saturable binding sites (K_D = 1 µM), numbering 8.6 × 10⁵ sites/cell. Both Hst 3 and Hst 4 competed for these binding sites with similar affinities, which is consistent with the micromolar concentration of Hsts required for candidacidal activity. Specific ¹²⁵I-Hst 5 binding was not detected to C. albicans spheroplasts, which were 14-fold less susceptible to Hst 5 killing, compared with intact cells in candidacidal assays. In overlay experiments, ¹²⁵I-Hst 5 bound to a 67-kDa protein detected in C. albicans whole cell lysates and crude membrane fractions, but not in the yeast cell wall fraction. Consistent with the overlay data, cross-linking of ¹²⁵I-Hst 5 to C. albicans resulted in the appearance of a specific 73-kDa ¹²⁵I-Hst 5-containing complex that was not detected in the cell wall. ¹²⁵I-Hst 5-binding protein of similar size was also observed in susceptible S. cerevisiae strain TI#20. This is the first description of Hst 5 binding sites on C. albicans which mediate cell killing and identification of a 67-kDa yeast Hst 5-binding protein. The binding characteristics of Hst 5 are in agreement with the observed potency of its biological effect and provide crucial information to the use of Hst 5 as a therapeutic agent. The presence of a specific C. albicans Hst 5-binding protein provides further insight into the potential mechanism of yeast killing and suggests a basis for differential activity between yeast killing and the nontoxic nature of Hsts to humans.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				W. L. Chaffin Candida albicans Cell Wall Proteins Microbiol. Mol. Biol. Rev., September 1, 2008; 72(3): 495 - 544. [Abstract] [Full Text] [PDF]

				W. S. Jang, X. S. Li, J. N. Sun, and M. Edgerton The P-113 Fragment of Histatin 5 Requires a Specific Peptide Sequence for Intracellular Translocation in Candida albicans, Which Is Independent of Cell Wall Binding Antimicrob. Agents Chemother., February 1, 2008; 52(2): 497 - 504. [Abstract] [Full Text] [PDF]

				S. Vylkova, W. S. Jang, W. Li, N. Nayyar, and M. Edgerton Histatin 5 Initiates Osmotic Stress Response in Candida albicans via Activation of the Hog1 Mitogen-Activated Protein Kinase Pathway Eukaryot. Cell, October 1, 2007; 6(10): 1876 - 1888. [Abstract] [Full Text] [PDF]

				S. Vylkova, N. Nayyar, W. Li, and M. Edgerton Human {beta}-Defensins Kill Candida albicans in an Energy-Dependent and Salt-Sensitive Manner without Causing Membrane Disruption Antimicrob. Agents Chemother., January 1, 2007; 51(1): 154 - 161. [Abstract] [Full Text] [PDF]

				X. S. Li, J. N. Sun, K. Okamoto-Shibayama, and M. Edgerton Candida albicans Cell Wall Ssa Proteins Bind and Facilitate Import of Salivary Histatin 5 Required for Toxicity J. Biol. Chem., August 11, 2006; 281(32): 22453 - 22463. [Abstract] [Full Text] [PDF]

				G.-X. Wei, A. N. Campagna, and L. A. Bobek Effect of MUC7 peptides on the growth of bacteria and on Streptococcus mutans biofilm J. Antimicrob. Chemother., June 1, 2006; 57(6): 1100 - 1109. [Abstract] [Full Text] [PDF]

				S. Vylkova, X. S. Li, J. C. Berner, and M. Edgerton Distinct Antifungal Mechanisms: {beta}-Defensins Require Candida albicans Ssa1 Protein, while Trk1p Mediates Activity of Cysteine-Free Cationic Peptides Antimicrob. Agents Chemother., January 1, 2006; 50(1): 324 - 331. [Abstract] [Full Text] [PDF]

				D. Baev, A. Rivetta, S. Vylkova, J. N. Sun, G.-F. Zeng, C. L. Slayman, and M. Edgerton The TRK1 Potassium Transporter Is the Critical Effector for Killing of Candida albicans by the Cationic Protein, Histatin 5 J. Biol. Chem., December 31, 2004; 279(53): 55060 - 55072. [Abstract] [Full Text] [PDF]

				L. de Repentigny, D. Lewandowski, and P. Jolicoeur Immunopathogenesis of Oropharyngeal Candidiasis in Human Immunodeficiency Virus Infection Clin. Microbiol. Rev., October 1, 2004; 17(4): 729 - 759. [Abstract] [Full Text] [PDF]

				A. Lupetti, C. P. J. M. Brouwer, H. E. C. Dogterom-Ballering, S. Senesi, M. Campa, J. T. van Dissel, and P. H. Nibbering Release of calcium from intracellular stores and subsequent uptake by mitochondria are essential for the candidacidal activity of an N-terminal peptide of human lactoferrin J. Antimicrob. Chemother., September 1, 2004; 54(3): 603 - 608. [Abstract] [Full Text] [PDF]

				M. S. Akhtar, J. Iqbal, M. A. Khan, J. Irfanullah, M. Jehangir, B. Khan, I. ul-Haq, G. Muhammad, M. A. Nadeem, M. S. Afzal, et al. 99mTc-Labeled Antimicrobial Peptide Ubiquicidin (29-41) Accumulates Less in Escherichia coli Infection than in Staphlococcus aureus Infection J. Nucl. Med., May 1, 2004; 45(5): 849 - 856. [Abstract] [Full Text] [PDF]

				G.-X. Wei and L. A. Bobek In vitro synergic antifungal effect of MUC7 12-mer with histatin-5 12-mer or miconazole J. Antimicrob. Chemother., May 1, 2004; 53(5): 750 - 758. [Abstract] [Full Text] [PDF]

				M. Viejo-Diaz, M. T. Andres, and J. F. Fierro Modulation of In Vitro Fungicidal Activity of Human Lactoferrin against Candida albicans by Extracellular Cation Concentration and Target Cell Metabolic Activity Antimicrob. Agents Chemother., April 1, 2004; 48(4): 1242 - 1248. [Abstract] [Full Text] [PDF]

				D. Wunder, J. Dong, D. Baev, and M. Edgerton Human Salivary Histatin 5 Fungicidal Action Does Not Induce Programmed Cell Death Pathways in Candida albicans Antimicrob. Agents Chemother., January 1, 2004; 48(1): 110 - 115. [Abstract] [Full Text] [PDF]

				J. Dong, S. Vylkova, X.S. Li, and M. Edgerton Calcium Blocks Fungicidal Activity of Human Salivary Histatin 5 through Disruption of Binding with Candida albicans J. Dent. Res., September 1, 2003; 82(9): 748 - 752. [Abstract] [Full Text] [PDF]

				X. S. Li, M. S. Reddy, D. Baev, and M. Edgerton Candida albicans Ssa1/2p Is the Cell Envelope Binding Protein for Human Salivary Histatin 5 J. Biol. Chem., August 1, 2003; 278(31): 28553 - 28561. [Abstract] [Full Text] [PDF]

				D. Baev, A. Rivetta, X. S. Li, S. Vylkova, E. Bashi, C. L. Slayman, and M. Edgerton Killing of Candida albicans by Human Salivary Histatin 5 Is Modulated, but Not Determined, by the Potassium Channel TOK1 Infect. Immun., June 1, 2003; 71(6): 3251 - 3260. [Abstract] [Full Text] [PDF]

				M. R. Yeaman and N. Y. Yount Mechanisms of Antimicrobial Peptide Action and Resistance Pharmacol. Rev., March 1, 2003; 55(1): 27 - 55. [Abstract] [Full Text] [PDF]

				L. A. Bobek and H. Situ MUC7 20-Mer: Investigation of Antimicrobial Activity, Secondary Structure, and Possible Mechanism of Antifungal Action Antimicrob. Agents Chemother., February 1, 2003; 47(2): 643 - 652. [Abstract] [Full Text] [PDF]

				K. Hieshima, H. Ohtani, M. Shibano, D. Izawa, T. Nakayama, Y. Kawasaki, F. Shiba, M. Shiota, F. Katou, T. Saito, et al. CCL28 Has Dual Roles in Mucosal Immunity as a Chemokine with Broad-Spectrum Antimicrobial Activity J. Immunol., February 1, 2003; 170(3): 1452 - 1461. [Abstract] [Full Text] [PDF]

				D. H. Fitzgerald, D. C. Coleman, and B. C. O'Connell Susceptibility of Candida dubliniensis to Salivary Histatin 3 Antimicrob. Agents Chemother., January 1, 2003; 47(1): 70 - 76. [Abstract] [Full Text] [PDF]

				D. Baev, X. S. Li, J. Dong, P. Keng, and M. Edgerton Human Salivary Histatin 5 Causes Disordered Volume Regulation and Cell Cycle Arrest in Candida albicans Infect. Immun., September 1, 2002; 70(9): 4777 - 4784. [Abstract] [Full Text] [PDF]

				A. Lupetti, A. Paulusma-Annema, S. Senesi, M. Campa, J. T. van Dissel, and P. H. Nibbering Internal Thiols and Reactive Oxygen Species in Candidacidal Activity Exerted by an N-Terminal Peptide of Human Lactoferrin Antimicrob. Agents Chemother., June 1, 2002; 46(6): 1634 - 1639. [Abstract] [Full Text] [PDF]

				D. Baev, X. Li, and M. Edgerton Genetically engineered human salivary histatin genes are functional in Candida albicans: development of a new system for studying histatin candidacidal activity Microbiology, December 1, 2001; 147(12): 3323 - 3334. [Abstract] [Full Text] [PDF]

				E. J. Helmerhorst, R. F. Troxler, and F. G. Oppenheim The human salivary peptide histatin 5 exerts its antifungal activity through the formation of reactive oxygen species PNAS, November 15, 2001; (2001) 141366998. [Abstract] [Full Text] [PDF]

				A. Lupetti, A. Paulusma-Annema, M. M. Welling, S. Senesi, J. T. van Dissel, and P. H. Nibbering Candidacidal Activities of Human Lactoferrin Peptides Derived from the N Terminus Antimicrob. Agents Chemother., December 1, 2000; 44(12): 3257 - 3263. [Abstract] [Full Text]

				M. Edgerton, S. E. Koshlukova, M. W. B. Araujo, R. C. Patel, J. Dong, and J. A. Bruenn Salivary Histatin 5 and Human Neutrophil Defensin 1 Kill Candida albicans via Shared Pathways Antimicrob. Agents Chemother., December 1, 2000; 44(12): 3310 - 3316. [Abstract] [Full Text]

				S. E. Koshlukova, M. W. B. Araujo, D. Baev, and M. Edgerton Released ATP Is an Extracellular Cytotoxic Mediator in Salivary Histatin 5-Induced Killing of Candida albicans Infect. Immun., December 1, 2000; 68(12): 6848 - 6856. [Abstract] [Full Text] [PDF]

				C. Gyurko, U. Lendenmann, R. F. Troxler, and F. G. Oppenheim Candida albicans Mutants Deficient in Respiration Are Resistant to the Small Cationic Salivary Antimicrobial Peptide Histatin 5 Antimicrob. Agents Chemother., February 1, 2000; 44(2): 348 - 354. [Abstract] [Full Text]

				Y. Xu, I. Ambudkar, H. Yamagishi, W. Swaim, T. J. Walsh, and B. C. O'Connell Histatin 3-Mediated Killing of Candida albicans: Effect of Extracellular Salt Concentration on Binding and Internalization Antimicrob. Agents Chemother., September 1, 1999; 43(9): 2256 - 2262. [Abstract] [Full Text]

				S. E. Koshlukova, T. L. Lloyd, M. W. B. Araujo, and M. Edgerton Salivary Histatin 5 Induces Non-lytic Release of ATP from Candida albicans Leading to Cell Death J. Biol. Chem., July 2, 1999; 274(27): 18872 - 18879. [Abstract] [Full Text] [PDF]

				E. J. Helmerhorst, W. van't Hof, P. Breeuwer, E. C. I. Veerman, T. Abee, R. F. Troxler, A. V. N. Amerongen, and F. G. Oppenheim Characterization of Histatin 5 with Respect to Amphipathicity, Hydrophobicity, and Effects on Cell and Mitochondrial Membrane Integrity Excludes a Candidacidal Mechanism of Pore Formation J. Biol. Chem., February 16, 2001; 276(8): 5643 - 5649. [Abstract] [Full Text] [PDF]

				E. J. Helmerhorst, R. F. Troxler, and F. G. Oppenheim The human salivary peptide histatin 5 exerts its antifungal activity through the formation of reactive oxygen species PNAS, December 4, 2001; 98(25): 14637 - 14642. [Abstract] [Full Text] [PDF]