Primary Structure and Anticandidal Activity of the Major Histatin from Parotid Secretion of the Subhuman Primate,Macaca fascicularis

Abstract

A major macaque histatin (M-histatin 1) from the parotid secretion of the subhuman primate, Macaca fascicularis, was isolated by gel filtration on Bio-Gel P-2 and purified to homogeneity by reversed-phase high-performance liquid chromatography on a TSK-ODS C₁₈ column. The complete amino acid sequence of M-histatin 1, determined by automated Edman degradation, is: 1 10 20 Asp-Pse-His-Glu-Glu-Arg-His-His-Gly-Arg-His-Gly-His-His-Lys-Tyr-Gly-Arg-Lys-Phe 21 30 38 His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr-Arg-Ser-Asn-Tyr-Leu-Tyr-Asp-Asn M-histatin 1 contains 38 amino acid residues, a phosphoserine at residue 2, has a molecular weight of 4881.8, a calculated pI of 8.5, and histidine forms 26.3% of the mass. The hydropathicity plot of M-histatin 1 predicts that the molecule is entirely hydrophilic, and Chou-Fasman secondary prediction indicates that the polypeptide is devoid of alpha-helix and beta-sheet conformation in aqueous solutions but contains a series of beta turns. M-histatin 1 includes a six-amino-acid insert (residue 10-15) not present in human histatins and, with the introduction of gaps to maximize homology, it displays 89% and 91% sequence similarity with human histatins 1 and 3, respectively. M-histatin 1 exhibited fungicidal and fungistatic effects against the dimorphic pathogen, Candida albicans, in three separate bioassays. Its anticandidal effects were comparable with or greater than those of human histatins 1, 3, and 5. M-histatins 2, 3, and 4 were not sequenced directly because insufficient materials were available, but the amino acid composition of M-histatin 3 was nearly identical to that of the N-terminal 20 amino acid residues of M-histatin 1. There appears to be only one major histatin in macaque parotid secretion in contrast to the family of histatins in human parotid and submandibular secretions, and the significance of this in the context of evolution and mechanism of action in anticandidal assays is discussed.

Get full access to this article

View all access options for this article.

References

Atkinson, J.C. ; Yeh, C. ; Oppenheim, F.G. ; Bermudez, D. ; Baum, B.J. ; and Fox, P.C. (1990): Elevation of Salivary Antimicrobial Proteins Following HIV-1 Infection, J AIDS 3 :41-48.

Austwick, P.K.G. ; Pepin, G. ; Thomson, J.C. ; and Yarrow, D. (1966): Candida albicans and Other Yeasts Associated with Animal Disease. In: Symposium on Candida Infections, H.I. Winner and R. Hurley , Eds., Edinburgh: Livingstone , pp. 89-100.

Azen, E.A. (1973): Properties of Salivary Basic Proteins Showing Polymorphism, Biochem Genet 9:69-86.

Baum, B.J. ; Bird, J.L. ; and Longton, R.W. (1977): Polyacrylamide Gel Electrophoresis of Human Salivary Histidine-rich Polypeptides, J Dent Res 56:1115-1118.

Bowen, W.H. (1981): Scoring Caries in Primates. In: Proceedings, Animal Models in Cariology, J.M. Tanzer , Ed., Sp. Supp. Microbiology Abstracts, Washington, D.C.: Information Retrieval, Inc., pp. 183-188.

Budtz-Jorgensen, E. (1975): Effects of Triamcinolone Acetonide on Experimental Oral Candidiasis in Monkeys, Scand J Dent Res 83:171-178.

Chou, P.Y. and Fasman, G.D. (1979): Prediction of the Secondary Structure of Proteins from their Amino Acid Sequence, Adv Enzymol Relat Areas Mol Biol 47:45-148.

Govindarajulu, Z. (1988): Statistical Techniques in Bioassay, Basel: Karger, pp. 28-64.

Gow, N.A.R. and Gooday, G.W. (1987): Cytological Aspects of Dimorphism in Candida albicans , CRC Crit Rev Microbiol 15 :73-78.

10.

Hay, D.I. ; Bennick, A. ; Schlesinger, D.H. ; Minaguchi, K. ; Madapallimattam, G. ; and Schluckebier, S.K. (1988): The Primary Structure of Six Human Salivary Acidic Proline-rich Proteins (PRP-1, PRP-2, PRP-3, PRP-4, PIF-s and PIFf), Biochem J 255:15-21.

11.

Hay, D.I. ; Moreno, E.C. ; and Schlesinger, D.H. (1979): Phosphoprotein-inhibitors of Calcium Phosphate Precipitation from Salivary Secretions, Inorg Persp Biol Med 2:271-285.

12.

Hay, D.I. ; Smith, D.J. ; Schluckebier, S.K. ; and Moreno, E.C. (1984): Relationship between Concentration of Human Salivary Statherin and Inhibition of Calcium Phosphate Precipitation in Stimulated Human Parotid Saliva, J Dent Res 63:857-863.

13.

Holbrook, I.B. and Molan, P.C. (1975): The Identification of a Peptide in Human Parotid Saliva Particularly Active in Enhancing the Glycolytic Activity of the Salivary Micro-organism, Biochem J 149:489-492.

14.

Kaufmann, A.F. and Quist, K.D. (1969): Thrush in a Rhesus Monkey: Report of a Case, Lab Anim Care 19:526-527.

15.

Kornman, K.S. ; Holt, S.C. ; and Robertson, P.B. (1981): The Microbiology of Ligature-induced Periodontitis in the Cynomolgus Monkey, J Periodont Res 16:363-371.

16.

Kyte, J. and Doolittle, R.F. (1982): A Simple Method for Displaying the Hydropathic Character of a Protein, J Mol Biol 157:105-132.

17.

Levitz, S.M. and Diamond, R.D. (1985): A Rapid Colorimetric Assay of Fungal Viability with the Tetrazolium Salt MTT, J Infect Dis 152:938-945.

18.

MacKay, B.J. ; Denepitiya, L. ; Iacono, V.J. ; Krost, S.P. ; and Pollock, J.J. (1984): Growth Inhibitory and Bactericidal Effects of Human Parotid Salivary Histidine-rich Polypeptides on Streptococcus mutans , Infect Immun 44:695-701.

19.

Minaguchi, K. and Bennick, A. (1989a): Genetics of Human Salivary Proteins, J Dent Res 68:2-15.

20.

Odds, F.H. (1979): Candida and Candidiasis, Leicester : Leicester University Press, pp. 29-31.

21.

Odds, F.H. (1980): A Simple System for the Presumptive Identification of Candida albicans and Differentiation of Strains within the Species, Sabouraudia 18:301-317.

22.

Oppenheim, F.G. (1989): Salivary Histidine-rich Proteins. In: Human Saliva: Clinical Chemistry and Microbiology, Vol. 1, J.O. Tenovuo , Ed., Florida: CRC Press, Inc., pp. 151-160.

23.

Oppenheim, F.G. ; Offner, G.D. ; and Troxler, R.F. (1982): Phosphoproteins in the Parotid Saliva from the Subhuman Primate Macaca fascicularis, J Biol Chem 257:9271-9282.

24.

Oppenheim, F.G. ; Offner, G.D. ; and Troxler, R.F. (1985): Amino Acid Sequence of a Proline-rich Phosphoglycoprotein from Parotid Secretion of the Subhuman Primate Macaca fascicularis, J Biol Chem 260:10671-10679.

25.

Oppenheim, F.G. ; Xu, T. ; McMillian, F.M. ; Levitz, S.M. ; Diamond, R.D. ; Offner, G.D. ; and Troxler, R.F. (1988): Histatins, a Novel Family of Histidine-rich Proteins in Human Parotid Secretion. Isolation, Characterization , Primary Structure and Fungistatic Effects on Candida albicans, J Biol Chem 263 :7472-7477.

26.

Oppenheim, F.G. ; Yang, Y. ; Diamond, R.D. ; Hyslop, D. ; Offner, G.D. ; and Troxler, R.F. (1986): The Primary Structure and Functional Characterization of the Neutral Histidine-rich Polypeptide from Human Parotid Saliva, J Biol Chem 261 :1177-1182.

27.

Peters, E.H. ; Goodfriend, T. ; and Azen, E.A. (1977): Human Pb, Human Post-Pb, and Nonhuman Primate Pb Proteins: Immunological and Biochemical Relationships, Biochem Genet 15:947-962.

28.

Pollock, J.J. ; Denepitiya, L. ; MacKay, B.J. ; and Iacono, V. (1984): Fungistatic and Fungicidal Activity of Human Parotid Salivary Histidine-rich Polypeptides on Candida albicans, Infect Immun 44:702-707.

29.

Raj, P.A. ; Edgerton, M.E. ; and Levine, M.J. (1990): Salivary Histatin 5: Dependence of Sequence, Chain Length, and Helical Conformation for Candidacidal Activity, J Biol Chem 265 :3898-3905.

30.

Santarpia, R.P., III ; Brant, E.C. ; Lai, K. ; Brasseur, M.M. ; Hong, A.L. ; and Pollock, J.J. (1988): A Comparison of the Inhibition of Blastospore Viability and Germ Tube Development in Candida albicans by Histidine Peptides and Ketoconazole, Arch Oral Biol 33:567-573.

31.

Schlesinger, D.H. and Hay, D.I. (1977): Complete Covalent Structure of Statherin, a Tyrosine-rich Acidic Peptide, which Inhibits Calcium Phosphate Precipitation from Human Parotid Saliva, J Biol Chem 252:1689-1695.

32.

Schlesinger, D.H. ; Hay, D.I. ; and Levine, M.J. (1989): Complete Primary Structure of Statherin, a Potent Inhibitor of Calcium Phosphate Precipitation, from the Saliva of the Monkey, Macaca arctoides, Int J Peptide Protein Res 34:374-380.

33.

Svanborg, A. and Svannerholm, L. (1961): Plasma Total Lipid, Cholesterol, Triglycerides , Phospholipids and Free Fatty Acids in a Healthy Scandinavian Population, Acta Med Scand 169:43-49.

34.

Troxler, R.F. ; Offner, G.D. ; Xu, T. ; Vanderspek, J.C. ; and Oppenheim, F.G. (1990): Structural Relationship between Human Salivary Histatins, J Dent Res 69:2-6.

35.

Xu, L. ; Fischer, T. ; and Pollock, J.J. (1989): Sequence Determination of Low Molecular Weight Salivary Histidine-rich Polypeptides from Electroblots, Peptide Res 2:373-375.

36.

Xu, T. and Oppenheim, F.G. (1989): Further Characterization of Anti-Candida Activities of Histatins, J Dent Res 68:405, Abst. No. 1787.

37.

Xu, T. and Oppenheim, F.G. (1990): Antibacterial Effects of Histatins on Different Mutans Streptococci Serotypes, J Dent Res 69:239, Abst. No. 1042.