Determination of the human salivary peptides histatins 1, 3, 5 and statherin by high-performance liquid chromatography and by diode-array detection
ABSTRACT A reversed-phase high-performance liquid chromatography (HPLC) method with diode-array detection for the quantification of several human salivary peptides is described. Sample pretreatment consisted of the acidification of whole saliva by phosphate buffer. This treatment produced precipitation of mucins, alpha-amylases and other high-molecular-mass salivary proteins, simultaneous inhibition of intrinsic protease activities and reduction of sample viscosity. Direct HPLC analysis by diode-array detection of the resulting acidic sample allowed one to quantify histatin 1, histatin 3, histatin 5, statherin, as well as uric acid, in normal subjects. Moreover, the groups of peaks pertaining to proline-rich proteins and cystatins were tentatively identified. The method can be useful in assessing the concentration of salivary peptides from normal subjects and from patients suffering oral and/or periodontal diseases.
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ABSTRACT: In recent years interest in the characterization of the human salivary proteome has increased in order to explore its diagnostic potential. Major constituents of human saliva are highly polymorphic proteins that may have biological roles in oral lubrication and protection, e.g. proline-rich proteins (PRPs), statherins, histatins and cystatins. Interestingly, many of theses proteins are rapidly degraded in the oral cavity by host-and bacteria-or viral-derived proteases. Thus, compre-hensive analysis of peptides up to 15 kDa (peptidomics) of whole saliva may yield basic information on proteolytic pat-terns. By employing a LC-ESI-MS/MS approach a total of 107 native peptides from 17 distinct protein precursors was identified from whole saliva. Subsequently the catalog of peptides was used to analyze inter-individual differences in sa-liva samples from four donors by differential peptide display technology. Genetic polymorphisms were found in peptides from the PRB4M_HUMAN and PROL4_HUMAN precursors. Analysis of the proposed N-and C-termini of the peptides revealed frequent cleavage after Lys or Arg which is characteristic for salivary kallikrein enzymes. Furthermore, we high-light the cleavage motif Gln/Gly in the PRP-C precursor, which suggests a new proteolytic pattern in saliva.The Open Proteomics Journal 10/2008; 1(1). DOI:10.2174/1875039700801010099
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ABSTRACT: An amperometric biosensor for monitoring the level of saliva protein histatin is described. The biosensor-sensing elements comprise a layer of salivary histatin antibody self-assembled onto a gold quartz crystal by covalent attachment. The biospecific interaction between the immobilized histatin antibody and histatin antigen was monitored via a novel electroactive indicator, silver monodispersed on the gold surface. The silver is oxidized or reduced and the resulting peak currents provide analytical information about the concentrations of the salivary proteins. A detection limit of 0.626 pg/ml was obtained.Sensor Letters 05/2005; 3(2):161-163. DOI:10.1166/sl.2005.024 · 0.56 Impact Factor
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ABSTRACT: The circadian rhythms of histatins 1, 3, 5, of statherin and uric acid were investigated in whole human saliva. Histatins showed a rhythm approximately synchronous with salivary flow rate (acrophase around 5 pm), the higher amplitude pertaining to histatin 1 (about 50% of the mesor). Uric acid showed a large rhythm asynchronous with flow rate and histatin concentrations (4.4 ± 1.4 am). Statherin did not show a significant circadian rhythm on five of six volunteers. This finding confirms that the secretion route of statherin is different from that of histatins.Biological Rhythm Research 04/2002; 33(2):213-222. DOI:10.1076/brhm.18.104.22.1684 · 1.22 Impact Factor