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J Dent Res 82(9): 753-757, 2003
© 2003 International and American Associations for Dental Research

RESEARCH REPORT
Biological

Degradation of Antimicrobial Histatin-variant Peptides in Staphylococcus aureus and Streptococcus mutans

J. Groenink*, A.L.A. Ruissen, D. Lowies, W. van ’t Hof, E.C.I. Veerman, and A.V. Nieuw Amerongen

Department of Dental Basic Sciences, Section of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), Van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands;

* corresponding author, j.groenink.obc.acta{at}med.vu.nl


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Histidine-free variants of salivary histatin 5 have a broad antimicrobial activity against various bacteria. In relation to a possible therapeutic application, we were interested in the susceptibility of these small peptides (14 amino acids long) to microbial proteinases and whether this affects their antimicrobial activity. Analyses by SDS-PAGE of supernatants of peptide-bacteria incubation showed a reduction in protein bands within 15 minutes’ incubation, as a result of cellular internalization. Degradation products of dhvar1 and dhvar2 appeared within one hour in the supernatants of Streptococcus mutans and Staphylococcus aureus. In contrast, the variants dhvar3 and dhvar4 were more resistant to degradation under the same conditions. MALDI-TOF analyses identified cleavage of dhvar1 and dhvar2 at Glu6. The N-terminal peptide part (1–6) of dhvar1 and 2 showed no bactericidal activity, while peptide fragment (7–14) showed a highly reduced bactericidal activity.

KEY WORDS: antimicrobial peptides • histatin • oral bacteria • proteinase


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
In human saliva, a variety of proteins is present with bactericidal or bacteriostatic properties (Scott and Hancock, 2000; Van ’t Hof et al., 2001). Among these, a small group of histidine-rich peptides has been identified and classified as histatins (Pollock et al., 1984; Oppenheim et al., 1988; Troxler et al., 1990). Histatins exhibit killing activities against numerous oral bacteria, among which are Streptococcus mutans and Porphyromonas gingivalis (MacKay et al., 1984; Murakami et al., 1991; Payne et al., 1991). In addition, they show anti-fungal activity against Candida albicans, an opportunistic yeast that causes infection, e.g., in immunocompromised patients (Atkinson et al., 1990; Pollock et al., 1992) and patients with a reduced saliva output (Arendorf and Walker, 1979; Mandel et al., 1992).

By replacing multiple amino acids in a putative active domain of histatin 5 (dh-5, residues 11–24), we have designed several peptides (dhvar1–4) with increased anti-Candida activity, even against amphotericin B- and fluconazole-resistant strains (Helmerhorst et al., 1997, 1999). The peptides also have a broad activity against various bacteria (Helmerhorst et al., 1997; Groenink et al., 1999), including methicillin-resistant Staphylococcus aureus (MRSA). Because of their small size and chemical structure, these peptides should be susceptible to proteinases, as has been found for histatins (Oppenheim et al., 1988; Xu et al., 1993). In a previous study, we found that the proteinase inhibitor cystatin, another potential contributor to innate immunity in saliva, is partially degraded by proteinases secreted by the oral pathogen P. gingivalis. Cleavage was determined at only one distinct site without affecting the cystatin activity (Blankenvoorde et al., 1996). In the present study, we examined for different bacteria whether they possess proteolytic activity that may function as a protective mechanism by affecting the antimicrobial activity of histatin 5 and its variant peptides.


   MATERIALS & METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Peptides
Histatin 5 (DSHAKRHHGYKRKFHEKHHSHRGY), variant peptides dhvar1 (KRLFKELKFSLRKY), dhvar2 (KRLFKELLFSLRKY), dhvar3 (KRLFKKLKFSLRKY), dhvar4 (KRLFKKLLFSLRKY), dhvar1 (1–6) (KRLFKE), dhvar1 (7–14) (LKFSLRKY), and dhvar2 (7–14) (LLFSLRKY), and a peptide fragment of salivary protein cystatin S (aa 81–95: LDTCAFHEQPELQKK) were prepared by solid-phase peptide synthesis as described previously (Van ’t Hof et al., 1991). All peptides showed a purity > 90%, as analyzed by reversed-phase HPLC (Helmerhorst et al., 1997). Peptides were labeled with FITC to establish cellular association by fluorescence microscopy, as described previously (Ruissen et al., 2001).

Strains and Growth Conditions
All bacteria were cultured on 5% horse blood agar plates (Oxoid no. 2, Basingstoke, UK) supplemented with hemin (5 mg/L) and menadione (1 mg/L). S. aureus (HG386) and Pseudomonas aeruginosa were grown aerobically for 24 hrs at 37°C. P. gingivalis (HG66) was grown anaerobically for 4 days at 37°C. Actinobacillus actinomycetemcomitans (Y4) and S. mutans (Ingbritt) were cultured in air plus 5% CO2 for 48 hrs at 37°C. For degradation experiments, additional strains of S. mutans (HG982 and NG8) and S. aureus (ATCC10832) were examined. Bacteria were harvested by being wiped from the agar plates with sterile cotton swabs and were transferred to PBS (10 mM potassium phosphate buffer, containing 150 mM sodium chloride, pH 7.4).

Determination of Antibacterial Activity
Each bacterial strain was tested at least twice in duplicate in killing assays, essentially as described previously (Groenink et al., 1999). In short, 50 µL of bacteria suspensions (3.2 x 106 cells/mL PBS) were incubated with equal volumes of serial dilutions of peptides (concentrations: 0.1–200 µg/mL PBS) at 37°C for 60 min. The incubation mixtures were diluted 200-fold in PBS, and 50-µL aliquots were plated onto blood agar plates so that cell viability could be determined. From the dose-response curves, the LC50-values, the peptide concentrations at which 50% of cells were killed, were read.

In addition to using the aforementioned conditions, we also performed assays with histatin 5 and control peptide cystatin S (81–95) in a 1-mM potassium phosphate buffer, pH 7.0 (PPB), since histatin 5 exerts appreciable microbicidal activity only in low-ionic-strength buffers (Helmerhorst et al., 1997; Ruissen et al., 2001).

Peptide Degradation
Degradation of 200 µg/mL of the peptides by 6.4 x 108 cells/mL at 37°C was monitored over a four-hour period. Samples were collected at t = 15, 60, 120, and 240 min, centrifuged twice at 10,000 g for 5 min (MSE Micro Centaur bench top centrifuge; Sanyo Gallenkamp, Loughborough, UK). Supernatants were heated (5 min at 100°C) to remove proteolytic activity.

To establish the different classes of enzymes involved in the peptide breakdown, we performed incubations of peptide and bacteria in the presence of 1 mM of the following proteolytic inhibitors: EDTA, PMSF, pepstatin A, E64, and bestatine. To investigate whether the different bacteria secrete enzymes capable of peptide-degradation, we grew cells in Brain Heart Infusion Medium (Oxoid, Basingstoke, UK). After 6 days, culture supernatants were isolated and incubated with dhvar1 solutions (200 µg/mL) in a ratio of 1:4 at 37°C for different time intervals (5, 15, 30, 60, 90 min).

The appearance of degradation products was analyzed by SDS-PAGE, for which samples were dissolved in reducing sample buffer (50 mM Tris buffer, pH 6.8, containing 2% SDS and 10 mM DTT) and heated for 5 min at 100°C. SDS-PAGE was performed on a Pharmacia PhastSystem (Pharmacia, Uppsala, Sweden) with high-density SDS gels. Protein bands were stained with 0.1% Coomassie Brilliant R 350 in 30% (v/v) methanol and 10% (v/v) acetic acid. Degradation products obtained were identified by means of MALDI-TOF (custom-built device).


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Killing Activity
Based on an active domain of histatin 5 (amino acids 11–24), 4 peptides (dhvar1, 2, 3, and 4) have been synthesized. Killing activities of histatin 5 and it variants were tested in a killing assay against various micro-organisms. The bactericidal properties of the peptides varied, depending on the micro-organism (TableGo)—e.g., dhvar1 exhibited a weaker cidal activity toward the Gram-negative A. actinomycetemcomitans (LC50 of 45.9 µM) than dhvar4 (LC50 of 5.2 µM), but comparable activities against the Gram-positive S. aureus and Gram-negative P. aeruginosa (see TableGo). For A. actinomycetemcomitans, S. aureus, S. mutans, and P. aeruginosa, the peptides dhvar2 and dhvar3 were less active than dhvar4, which overall was the most active peptide. The Gram-negative organism P. gingivalis was equally susceptible to all 4 peptides (LC50 values between 4.3 and 5.2 µM).


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  		Table. LC50 Values (µM) of Histatin 5 and Dhvar Peptides against Several Bacteria
 
At elevated ionic strength (PBS), none of the bacteria tested was killed by histatin 5, except for P. aeruginosa, for which an LC50 value of 24.1 µM was found. In contrast, in PPB (1 mM potassium phosphate buffer), histatin 5 exhibited high bactericidal activities (LC50 <= 1.7 µM). Of P. aeruginosa, 95% of the bacteria were killed at histatin 5 concentrations of 0.1 µM. Control peptide cystatin S (81–95) showed no bactericidal activity against any of the bacteria, either in PBS or in PPB.

Degradation of Antimicrobial Peptides
Supernatants of peptide-bacteria mixtures were analyzed on high-density SDS-gels. After 1 hr of incubation, the intensity of the dhvar bands was decreased (Fig. 1Go, lane 2). A decrease in the histatin 5 band was observed only when the incubation was conducted in PPB. Fluorescence microscopy with FITC-labeled peptides showed homogeneous intracellular distribution of the label (not shown), suggesting that peptides were absorbed from the supernatant by internalization. No bacteria were fluorescent when incubated with FITC-labeled cystatin S (81–95).



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  		Figure 1. Peptide degradation by bacterial proteinases. (A) S. aureus cells (6.4 x 108 cells/mL PBS) were incubated with the dhvar peptides for 4 hrs at 37°C. Samples were analyzed on high-density SDS-PAGE gels in combination with Coomassie Brilliant Blue staining. Lane 1, dhvar1 (200 µg/mL) not incubated with bacteria; lanes 2–4, samples taken at 15, 120, and 240 min, respectively. A peptide band with lower molecular weight gradually appeared over time, indicating progressive degradation. Incubation of S. mutans with dhvar1 and dhvar2 showed similar patterns (not shown). (B) Lane 1 contained the native peptide dhvar1 (200 µg/mL). Lane 2: Dhvar1 incubated with P. gingivalis cells (1.6 x 106 cells/mL PBS) for 60 min at 37°C. Lane 3: Dhvar1 incubated (4:1) with supernatant of 6 days’ cultured P. gingivalis (five-minute incubation). Incubations with P. aeruginosa showed similar results (not shown).

 
Incubations of dhvar1 and dhvar2 with S. mutans (Ingbritt) or S. aureus (HG386) cells showed the appearance of new protein bands, migrating to a lower molecular position in the SDS gels. Sampling at different time points showed a progressive decrease of the original material. Concomitantly, new protein bands, most likely degradation products, appeared (compare Fig. 1AGo, lane 2 [15 min] and lane 4 [240 min]). Additional incubations of dhvar1 with different strains of S. mutans (HG982 and NG8) and S. aureus (ATCC10832) showed similar patterns of peptide degradation. Incubations of the different S. mutans strains with either dhvar3 or dhvar4 did not result in the appearance of putative breakdown products in the supernatant (not shown). No breakdown was observed for peptides dhvar3 and dhvar4 when incubated with S. aureus strain HG386, while degradation seemed to occur following incubations with S. aureus strain ATCC10832.

For P. gingivalis and P. aeruginosa, two pathogens known to possess a variety of proteolytic enzymes, uptake of peptide from the supernatant was observed, but no breakdown products were detected by SDS-PAGE (Fig. 1BGo, lane 2). In contrast, incubations of dhvar1 and dhvar4 with six-day culture supernatant of these bacteria showed peptide degradation within 5 min of incubation (Fig. 1BGo, lane 3), indicating the bacteria secreted peptide-degrading enzymes. No peptide degradation was observed when peptides were incubated with either cells and culture supernatants of A. actinomycetemcomitans or culture supernatants of S. mutans and S. aureus.

Identification of Breakdown Products and Their Antimicrobial Activity
The peptides dhvar1 and dhvar2 differ from dhvar3 and dhvar4, respectively, in one amino acid (Glu6 instead of Lys6). A serine proteinase of S. aureus, referred to as V8 proteinase, is a known endoproteinase for hydrolysis after a glutamate residue (Houmard and Drapeau, 1972). Electrophoretic analyses of dhvar peptides incubated with V8 proteinase (10 µg/mL, Boehringer Mannheim, Germany) demonstrated degradation of dhvar1 and dhvar2 within 5 min (not shown), while dhvar3 and dhvar4 were resistant to overnight incubations with V8. The cystatin peptide, containing a glutamate residue, was hydrolyzed by V8 proteinase, but not when incubated with S. aureus and S. mutans cells. Taken together, these experiments suggest that the peptide degradation by S. aureus and S. mutans is an intracellular process.

To obtain indications of the enzymes involved, we incubated peptides dhvar1 and dhvar2 with S. mutans in the presence of specific inhibitors of proteolytic enzymes. It appeared that peptide degradation was inhibited by pepstatin, an inhibitor of aspartic proteinases (not shown). Other inhibitors tested—including PMSF (serine proteinases), EDTA (metalloproteinases), E64 (cysteine proteinases), and bestatin (aminopeptidases)—had no effect.

Fig. 2Go shows a mass spectrum of the incubation supernatant of dhvar1, obtained after 5 hours’ incubation with S. mutans. The MALDI-TOF profile confirmed that breakdown products of dhvar1 were released in the incubation supernatant, and new peaks could be assigned to peptide fragments KRLFKE [dhvar1 (1–6); Mw 820] and LKFSLRKY [dhvar1 (7–14); Mw 1054]. We synthesized cleavage products of dhvar1 and dhvar2 to test whether digestion impaired the bactericidal activities toward S. aureus and S. mutans.



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  		Figure 2. Degradation of dhvar1 by S. mutans analyzed by MALDI-TOF. A suspension of S. mutans (6.4 x 108 cells/mL PBS) was incubated with 200 µg/mL dhvar1 for 5 hrs at 37°C. Subsequently, cells were removed by centrifugation, proteinases were inactivated by being heated for 5 min at 100°C, and samples were analyzed by MALDI-TOF. The original peptide (molecular mass 1855) was degraded into two fragments with molecular masses of 821 and 1054, corresponding to the fragments Lys1-Glu6 and Leu7-Tyr14, respectively. Potassium and sodium adducts contribute to minor adjacent peaks.

 
No bactericidal activity could be found for the N-terminal fragment (1–6) at concentrations as high as 244 µM. The bactericidal activity of the dhvar1 (7–14) fragment was much weaker than that of the intact peptide, exhibiting LC50 values of 154 µM for S. aureus and 136 µM for S. mutans, compared with 8.8 and 1.7 µM, respectively, for the original peptide. For fragment dhvar2 (7–14), no killing of S. aureus was found at the highest concentration tested (192 µM), while for S. mutans, an LC50 value of 140 µM was found, compared with 8.5 µM for the intact peptide.

Although within the time frame of the killing assay, no breakdown products of histatin 5 were found, MALDI-TOF analyses of histatin 5, incubated for 5 hrs with S. aureus, showed that degradation had occurred at Glu16, His18, His19, Ser20, and His21. Although more putative cleavage sites (His and Ser) are present in histatin 5, no additional breakdown products were found, suggesting that degradation occurred gradually at the C-terminal end.


   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
In the development of new peptides for pharmaceutical purposes, we wondered whether bacteria are able to inactivate cationic antimicrobial peptides by degradation and whether new variants can be designed that are resistant to degradation. Previously, degradation of histatin 5 has been demonstrated for S. sanguis (Payne et al., 1991). In addition, we found that histatin 5, dh-5, and P-113, a peptide containing amino acid residues 4–15 of histatin 5, were susceptible to degradation by C. albicans after the peptides had crossed the plasma membrane (Ruissen et al., 2003). Sajjan et al.(2001) found that P-113 was not active in the presence of purulent sputum from cystic fibrosis patients. In contrast, the mirror image P-113D, which is resistant to proteolytic degradation due to the stereo-specificity of proteinases, was active in sputum against pathogens of cystic fibrosis, e.g., P. aeruginosa and S. aureus. Our results identified a gradual degradation of histatin 5 by S. aureus. In 5 hrs, breakdown products still contained the motif of P-113. Together, not only is the degradation of histatins in saliva ascribed to host proteinases (Oppenheim et al., 1988), but also microbial proteinases seem to contribute.

We have developed histidine-free variants of histatin 5 with bactericidal activities higher than those of histatin 5 (TableGo). Peptides dhvar1 and dhvar2 seemed susceptible to several strains of S. mutans and S. aureus. The digestive products showed low bactericidal activities toward these 2 bacteria. Although only one cleavage site was established, i.e., Glu6, the rate of degradation seemed to differ slightly for the 2 peptides, perhaps as a result of the variation at amino acid position 8 (Lys or Leu). Strikingly, dhvar1 and dhvar2 were degraded by extracellular proteinase V8 of S. aureus (Houmard and Drapeau, 1972), but no proteolytic activity was found in S. aureus culture supernatant. In this context, we did not verify, under our culturing conditions, whether the S. aureus strains released V8 proteinase into the culture medium.

In our experimental design, dhvar3 and dhvar4 seemed more resistant to degradation, since for only one S. aureus strain was degradation observed after cellular internalization. Nevertheless, culture supernatants of P. gingivalis and P. aeruginosa contained proteolytic activity toward both dhvar1 and dhvar4. Conclusively, degradation of antimicrobial peptides seemed to occur by both extracellular and, more to our interest, intracellular microbial proteinases. We expect that degradation prior to cellular uptake should affect the killing activity to a great extent, while we found that intracellular degradation occurred after killing was established. For pharmaceutical application, the use of mirror images of antimicrobial peptides (Sajjan et al., 2001) or peptide mixtures with a protective polymeric carrier, which has been demonstrated to reduce the susceptibility of dhvar peptides (Ruissen et al., 1999), should be considered.


   ACKNOWLEDGMENTS
 
The authors thank K. Nazmi of the Academic Centre for Dentistry Amsterdam for peptide synthesis, and Dr. R.C. van der Schors of the Vrije Universiteit in Amsterdam for mass spectrometry. This research is financially supported by the Dutch Technology Foundation, grant VTH3950, and by Sarah Lee/Douwe Egberts.

Received May 28, 2002; Last revision May 19, 2003; Accepted June 25, 2003


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Arendorf TM, Walker DM (1979). Oral candidal populations in health and disease. Br Dent J 147:267–272.[ISI][Medline]

Atkinson JC, Yeh C, Oppenheim FG, Bermudez D, Baum BJ, Fox PC (1990). Elevation of salivary antimicrobial proteins following HIV-1 infection. J Acquir Immune Defic Syndr 3:41–48.

Blankenvoorde MF, Henskens YM, van ’t Hof W, Veerman EC, Nieuw Amerongen AV (1996). Inhibition of the growth and cysteine proteinase activity of Porphyromonas gingivalis by human salivary cystatin S and chicken cystatin. Biol Chem 377:847–850.[ISI][Medline]

Groenink J, Walgreen-Weterings E, van ’t Hof W, Veerman EC, Nieuw Amerongen AV (1999). Cationic amphipathic peptides, derived from bovine and human lactoferrins, with antimicrobial activity against oral pathogens. FEMS Microbiol Lett 179:217–222.[ISI][Medline]

Helmerhorst EJ, van ’t Hof W, Veerman EC, Simoons-Smit I, Nieuw Amerongen AV (1997). Synthetic histatin analogues with broad-spectrum antimicrobial activity. Biochem J 326:39–45.

Helmerhorst EJ, Reijnders IM, van ’t Hof W, Simoons-Smit I, Veerman EC, Nieuw Amerongen AV (1999). Amphotericin B- and fluconazole-resistant Candida spp., Aspergillus fumigatus, and other newly emerging pathogenic fungi are susceptible to basic antifungal peptides. Antimicrob Agents Chemother 43:702–704.[Abstract/Free Full Text]

Houmard J, Drapeau GR (1972). Staphylococcal protease: a proteolytic enzyme specific for glutamoyl bonds. Proc Natl Acad Sci USA 69:3506–3509.[Abstract/Free Full Text]

MacKay BJ, Denepitiya L, Iacono VJ, Krost SB, Pollock JJ (1984). Growth-inhibitory and bactericidal effects of human parotid salivary histidine-rich polypeptides on Streptococcus mutans. Infect Immun 44:695–701.[Abstract/Free Full Text]

Mandel ID, Barr CE, Turgeon L (1992). Longitudinal study of parotid saliva in HIV-1 infection. J Oral Pathol Med 21:209–213.[ISI][Medline]

Murakami Y, Nagata H, Amano A, Takagaki M, Shizukuishi S, Tsunemitsu A, et al. (1991). Inhibitory effects of human salivary histatins and lysozyme on coaggregation between Porphyromonas gingivalis and Streptococcus mitis. Infect Immun 59:3284–3286.[Abstract/Free Full Text]

Oppenheim FG, Xu T, McMillian FM, Levitz SM, Diamond RD, Offner GD, et al. (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.[Abstract/Free Full Text]

Payne JB, Iacono VJ, Crawford IT, Lepre BM, Bernzweig E, Grossbard BL (1991). Selective effects of histidine-rich polypeptides on the aggregation and viability of Streptococcus mutans and Streptococcus sanguis. Oral Microbiol Immunol 6:169–176.[ISI][Medline]

Pollock JJ, Denepitiya L, MacKay BJ, Iacono VJ (1984). Fungistatic and fungicidal activity of human parotid salivary histidine-rich polypeptides on Candida albicans. Infect Immun 44:702–707.[Abstract/Free Full Text]

Pollock JJ, Santarpia RP 3rd, Heller HM, Xu L, Lal K, Fuhrer J, et al. (1992). Determination of salivary anticandidal activities in healthy adults and patients with AIDS: a pilot study. J Acquir Immune Defic Syndr 5:610–618.

Ruissen AL, van der Reijden WA, van ’t Hof W, Veerman EC, Nieuw Amerongen AV (1999). Evaluation of the use of xanthan as vehicle for cationic antifungal peptides. J Control Release 60:49–56.[ISI][Medline]

Ruissen AL, Groenink J, Helmerhorst EJ, Walgreen-Weterings E, van ’t Hof W, Veerman EC, et al. (2001). Effects of histatin 5 and derived peptides on Candida albicans. Biochem J 356:361–368.[ISI][Medline]

Ruissen AL, Groenink J, Krijtenberg P, Walgreen-Weterings E, van ’t Hof W, Veerman EC, et al. (2003). Internalisation and degradation of histatin 5 by Candida albicans. Biol Chem 384:183–190.[ISI][Medline]

Sajjan US, Tran LT, Sole N, Rovaldi C, Akiyama A, Friden PM, et al. (2001). P-113D, an antimicrobial peptide active against Pseudomonas aeruginosa, retains activity in the presence of sputum from cystic fibrosis patients. Antimicrob Agents Chemother 45:3437–3444.[Abstract/Free Full Text]

Scott MG, Hancock RE (2000). Cationic antimicrobial peptides and their multifunctional role in the immune system. Crit Rev Immunol 20:407–431.[ISI][Medline]

Troxler RF, Offner GD, Xu T, Vanderspek JC, Oppenheim FG (1990). Structural relationship between human salivary histatins. J Dent Res 69:2–6.[Abstract/Free Full Text]

Van ’t Hof W, Driedijk PC, van den Berg M, Beck-Sickinger AG, Jung G, Aalberse RC (1991). Epitope mapping of the Dermatophagoides pteronyssinus house dust mite major allergen Der p II using overlapping synthetic peptides. Mol Immunol 28:1225–1232.[ISI][Medline]

Van ’t Hof W, Veerman EC, Helmerhorst EJ, Amerongen AV (2001). Antimicrobial peptides: properties and applicability. Biol Chem 382:597–619.[ISI][Medline]

Xu L, Lal K, Santarpia RP 3rd, Pollock JJ (1993). Salivary proteolysis of histidine-rich polypeptides and the antifungal activity of peptide degradation products. Arch Oral Biol 38:277–283.[ISI][Medline]




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