Article

Determination of the human salivary peptides histatins 1, 3, 5 and statherin by high-performance liquid chromatography and by diode-array detection.

Department of Sciences Applied to Biosystems, Cagliari University, Italy.
Journal of chromatography. B, Biomedical sciences and applications 03/2001; 751(1):153-60. DOI: 10.1016/S0378-4347(00)00466-7
Source: PubMed

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.

0 Bookmarks
 · 
274 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: Interest in the characterization of the salivary proteome has increased in the last few years. This review discusses the different techniques and methodologies applied to the separation and identification of salivary proteins. Nowadays, proteomic techniques are the state of the art for the analysis of biologic materials and saliva is no exception. 2D electrophoresis and tryptic digest analysis by mass spectrometry are the typical methodology, but new approaches using 2D liquid chromatography/mass spectrometry methods have already been introduced for saliva analysis. Due to their important physiologic role in the oral cavity, low-molecular-weight proteins and peptides are also included in this article and the methodologies discussed.
    Expert Review of Proteomics 09/2005; 2(4):521-39. · 3.90 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Saliva is a glandular secretion that is vital in the maintenance of healthy oral tissues. In this review we outline the high abundance salivary proteins, summarise the status of the salivary proteome and peptidome, the genetic origin and recognised functions of these proteins, the diseases associated with salivary disorders, and the emerging saliva-derived peptide therapeutics. Different proteomic approaches have reported the identification of over 1,300 proteins in saliva. However there are fewer than 100 high abundance proteins, identified by multiple methods including, two-dimensional polyacrylamide gel electrophoresis and HPLC combined with mass spectrometry. Analysis of the genes coding for the salivary proteins demonstrated a non-uniform chromosomal distribution with chromosome 4 having the largest proportion of genes expressed in salivary glands. Several diseases are associated with salivary disorders including Sjögren’s syndrome, Prader-Willi syndrome, dental caries and stress related disorders. Saliva as a diagnostic medium for various biochemical tests has provided a non-invasive and accessibility advantage over other more regularly tested body fluids such as blood and urine. To-date the emerging saliva-based therapeutics include artificial salivas and antimicrobial agents based on histatins and mucins.
    International Journal of Peptide Research and Therapeutics 13(4):547-564. · 1.28 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Recently, we demonstrated that anti-statherin monoclonal antibodies could be generated upon immunisation of mice with in vivo formed human acquired enamel pellicle, indicating that statherin is a constituent of pellicle. To gain insight in the in vivo adsorption behaviour of statherin we tested the abundance of statherin in pellicle and investigated the relationship between statherin and protein levels in salivary secretions and pellicle using a capture ELISA. Statherin levels were approximately 20-fold higher in parotid and submandibular-sublingual secretions than in cleared whole saliva supernatant or pellicle, suggesting the rapid degradation of statherin in the oral cavity. A strong positive correlation was observed between statherin and protein levels in pellicle but not in saliva indicating that statherin and protein adsorption to pellicle are related processes. This indicates that statherin represents the integral part of proteins that constitute the pellicle structure and may play a key role in its formation.
    Archives of Oral Biology 06/2004; 49(5):379-85. · 1.55 Impact Factor