Polyacrylamide Gel Electrophoresis of Human Salivary Histidine-Rich-Polypeptides

Baum, Bruce J.; Bird, Janice L.; Longton, Robert W.

doi:10.1177/00220345770560091801

Cited by 40 publications

(34 citation statements)

References 11 publications

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“…Histatin 7 is in fact a mixture of small histatins (histatins 7-12) that has been resolved into six components (see below). The nomenclature used for histatins described in this paper is consistent with the original observations on the electrophoretic mobility of histatins in the cationic PAGE system (Baum et al, 1976(Baum et al, , 1977.…”

Section: Methodssupporting

confidence: 66%

“…-Histatins were examined on 15% slab gels with use of a cationic sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) system described previously (Baum et al, 1976(Baum et al, , 1977Oppenheim et al, 1988). The amino acid compositions of selected histatins and derived peptides were determined with use of a Beckman system 6300 amino acid analyzer following hydrolysis in 6 mol/L HCl for 22 h in vacuo at 1 100C.…”

Section: Methodsmentioning

confidence: 99%

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Structural Relationship Between Human Salivary Histatins

Troxler

Offner

Xu³

et al. 1990

J Dent Res

162

112

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Histatins are a group of electrophoretically distinct histidine-rich polypeptides with microbicidal activity found in human parotid and submandibular gland secretions. Recently, we have shown that histatins 1, 3, and 5 are homologous proteins that consist of 38, 32, and 24 amino acid residues, respectively, and that these polypeptides kill the pathogenic yeast, Candida albicans. We now describe the isolation and structural characterization of histatins 2, 4, 6, and 7-12, the remaining members ofthis group ofpolypeptides. Histatin 2 wasfound to be identical to the carboxyl terminal 26 residues of histatin 1; histatin 4 was found to be identical to the carboxyl terminal 20 residues of histatin 3; and histatin 6 was found to be identical to histatin 5, but contained an additional carboxyl terminal arginine residue. The amino acid sequences of histatins 7-12 formally correspond to residues 12-24, 13-24, 12-25, 13-25, 5-11, and 5-12, respectively, of histatin 3, but could also arise proteolytically from histatin 5 or 6. These results establish, for the first time, the complete structural relationships between all members of this group ofmicrobicidal proteins in human parotid saliva. The relationship of histatins to one another is discussed in the context of their genetic origin, biosynthesis and secretion into the oral cavity, and potential as reagents in anti-candidal studies.

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Section: Methodssupporting

confidence: 66%

Section: Methodsmentioning

confidence: 99%

Structural Relationship Between Human Salivary Histatins

Troxler

Offner

Xu³

et al. 1990

J Dent Res

162

112

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“…Tris-Tricine SDS-PAGE was performed as described in reference 26, with a separating gel containing 9.6% acrylamide, 0.3% bisacrylamide, and 3.6 M urea. Cationic PAGE was performed to separate the positively charged proteins and was carried out on a separating gel containing 15% acrylamide and 0.4% bisacrylamide (1,21).…”

Section: Methodsmentioning

confidence: 99%

Dialysis Unmasks the Fungicidal Properties of Glandular Salivary Secretions

et al. 2004

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Several salivary proteins exhibit fungicidal activity against the opportunistic oral pathogen Candida albicans when they are tested as pure proteins in vitro. However, salivary secretions that are examined by the same assays either lack or exhibit very low candidacidal activity. Since ionic strength is known to have an inhibitory effect on the fungicidal activities of some proteins, parotid secretion was subjected to dialysis with membranes having molecular weight cutoffs (MWCOs) of 500, 1,000, 10,000, and 25,000. Dialysis with membranes with MWCOs of >1,000 promoted fungicidal activity of parotid secretion, and this activity was dose dependent. The addition of sodium chloride to dialyzed, fungicidal parotid secretion abolished this activity, indicating that the fungicidal component was salt sensitive. Similar results were obtained with submandibular and sublingual secretions. Polyacrylamide gel electrophoresis under native and denaturing conditions was used to analyze the composition of the dialysate. Unexpectedly, proteins with MWs much lower than the nominal MWCOs of the membranes were not lost during dialysis. Among the retained proteins, the two fractions with MWs of approximately 17,000 and 4,000 exhibited fungicidal activity. These results are consistent with the presence of lysozyme and histatins, respectively, which may represent the major candidacidal capacity of dialyzed parotid secretion.

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“…In this system, histatins were resolved into Pb bands: a, b, c, d, and e. Using the same system, Peters et al (1977) resolved histatins into the same bands and one additional band, PPb. When five different electrophoresis systems were compared (Baum et al, 1977a), it was determined that a "very cationic" system of a 15% acrylamide gel, pH 2.7, gave the greatest resolution of histatin fractions from saliva, giving rise to histidine-rich proteins 1-7 (HRP-1-7). Later, researchers mainly followed the protocols of Baum et al (1977a), with some modifications (MacKay et al, 1984a;Oppenheim et al, 1988;Raj et al, 1990;Tsai et al, 1996).…”

Section: (I) Introductionmentioning

confidence: 99%

Human Salivary Histatins: Promising Anti-Fungal Therapeutic Agents

Tsai

Bobek

1998

Critical Reviews in Oral Biology & Medicine

141

115

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Histatins constitute a group of small, cationic multifunctional proteins present in the saliva of human and some nonhuman primates. The most significant function of histatins may be their anti-fungal activity against Candida albicans and Cryptococcus neoformans. Histatins have been extensively studied at both the protein and gene levels. The structure-function relationship of histatins with respect to their candidacidal activity has also been studied by means of recombinant histatin variants, as well as by chemically synthesized histatin fragments. The mechanism of histatins' action on Candida albicans is not clear, but it appears to be different from that of azole-based anti-fungal drugs which interrupt ergosterol synthesis. During the past 20 years, fungal infections have become more prevalent as a result of the emergence of AIDS, as well as, paradoxically, modern medical advances. The toxicity of current anti-fungal medicine, the emergence of drug-resistant strains, and the availability of only a few types of antifungal agents are the major disadvantages of current anti-fungal therapy. Therefore, the importance of the search for new, broadspectrum anti-fungals with little or no toxicity cannot be overemphasized. The following properties make histatins promising antifungal therapeutic agents: (1) They have little or no toxicity; (2) they possess high cidal activities against azole-resistant fungal species and most of the fungal species tested; and (3) their candidacidal activity is similar to that of azole-based antifungals.Current research efforts focus on the development of improved histatins with enhanced cidal activity and stability, and of suitable and effective histatin delivery systems. These and other approaches may help to outpace the growing list of drug-resistant and opportunistic fungi causing life-threatening, disseminating diseases. The histatins with improved protective properties may also be used as components of artificial saliva for patients with salivary dysfunction.

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Polyacrylamide Gel Electrophoresis of Human Salivary Histidine-Rich-Polypeptides

Cited by 40 publications

References 11 publications

Structural Relationship Between Human Salivary Histatins

Structural Relationship Between Human Salivary Histatins

Dialysis Unmasks the Fungicidal Properties of Glandular Salivary Secretions

Human Salivary Histatins: Promising Anti-Fungal Therapeutic Agents

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