scholarly journals The presence and origin of phosphopeptides in human saliva

1988 ◽  
Vol 250 (1) ◽  
pp. 171-177 ◽  
Author(s):  
K Minaguchi ◽  
G Madapallimattam ◽  
A Bennick
Keyword(s):  
Oral Cavity ◽  
Calcium Binding ◽  
Proteolytic Enzymes ◽  
Biological Activities ◽  
Human Saliva ◽  
Serine Proteinases ◽  
Rapid Degradation ◽  
Whole Saliva ◽  
Oral Surface ◽  
Hydroxyapatite Formation

The presence of phosphopeptides in whole saliva (saliva expectorated from the mouth) was demonstrated and their origin was evaluated. Whole saliva contained much larger numbers of small phosphopeptides than are found in the glandular secretions. Most of these originated from the acidic proline-rich proteins (PRPs) in the major salivary glands and were formed, after secretion into the oral cavity, as a result of rapid degradation by proteolytic enzymes from extraglandular sources contained in sediment from whole saliva. Some peptides may have been formed by cleavage of basic PRPs, but other phosphoproteins apparently contributed little to the observed phosphopeptides. Most of the enzymes that produced phosphopeptides are serine proteinases. The gel-electrophoretic band patterns of the phosphopeptides obtained from 26 individuals of various acidic-PRP phenotypes were remarkably similar, demonstrating that the enzymes responsible were generally present in the population surveyed and that similar cleavages occur regardless of the nature of the acidic PRPs. Many of these peptides were N-terminal proteolytic cleavage products of acidic PRPs. The N-terminal phosphorylated region of acidic PRPs contains various biological activities, such as inhibition of hydroxyapatite formation, calcium binding and binding to hydroxyapatite, the major mineral of teeth. The demonstration of these phosphopeptides in the saliva that is in contact with the oral surface may therefore be of biological importance.

scholarly journals Salivary protein roles in oral health and as predictors of caries risk

2018 ◽  
Vol 13 (1) ◽  
pp. 174-200
Author(s):  
Galina Laputková ◽  
Vladimíra Schwartzová ◽  
Juraj Bánovčin ◽  
Michal Alexovič ◽  
Ján Sabo
Keyword(s):  
Dental Caries ◽  
Oral Cavity ◽  
Human Saliva ◽  
Salivary Protein ◽  
Oral Diseases ◽  
Irreversible Damage ◽  
Current State ◽  
Whole Saliva ◽  
State Of Research ◽  
Insight Into

AbstractThis work describes the current state of research on the potential relationship between protein content in human saliva and dental caries, which remains among the most common oral diseases and causes irreversible damage in the oral cavity. An understanding the whole saliva proteome in the oral cavity could serve as a prerequisite to obtaining insight into the etiology of tooth decay at early stages. To date, however, there is no comprehensive evidence showing that salivary proteins could serve as potential indicators for the early diagnosis of the risk factors causing dental caries. Therefore, proteomics indicates the promising direction of future investigations of such factors, including diagnosis and thus prevention in dental therapy.

scholarly journals Identification and Characterization of a Nonimmunoglobulin Factor in Human Saliva That Inhibits Streptococcus mutans Glucosyltransferase

2002 ◽  
Vol 70 (3) ◽  
pp. 1136-1142 ◽  
Author(s):  
Christina Jespersgaard ◽  
George Hajishengallis ◽  
Michael W. Russell ◽  
Suzanne M. Michalek
Keyword(s):  
Molecular Weight ◽  
Cell Surface ◽  
Oral Cavity ◽  
High Molecular Weight ◽  
Streptococcus Mutans ◽  
Human Saliva ◽  
Size Exclusion ◽  
Enzyme Linked Immunosorbent Assay ◽  
Innate Defense ◽  
Whole Saliva

ABSTRACT Saliva contains an array of nonimmunoglobulin defense factors which are thought to contribute to the protection of the hard and soft tissue surfaces of the oral cavity by modulating microbial colonization and metabolism. Here we report the discovery of a putative innate defense factor in human saliva that inhibits the glucosyltransferase (GTF) of Streptococcus mutans, a virulence enzyme involved in oral colonization by this pathogen. The GTF-inhibiting factor (GIF) was initially identified as a nonimmunoglobulin salivary component that interfered with detection of antibodies to the glucan-binding region (GLU) of GTF by an enzyme-linked immunosorbent assay. This inhibitory activity was present in whole saliva and submandibular-sublingual saliva, but it was essentially absent from parotid saliva. GIF inhibited the recognition of S. mutans cell surface-associated GTF by specific antibodies but had no effect on antibodies to other cell surface antigens, suggesting that GIF specifically binds to GTF on S. mutans. GIF purified by size exclusion or affinity chromatography was used for biochemical and functional characterization. Analysis of GIF by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a high-molecular-weight glycoprotein after staining with Coomassie blue or Schiff's reagent. Heating and reduction with 2-mercaptoethanol of GIF resulted in the release of a ∼58-kDa protein that was identified as α-amylase by Western blotting using anti-α-amylase antibodies. GLU bound blotted α-amylase, suggesting that the latter molecule is the GLU-binding component of the GIF complex. The ability of GTF to synthesize extracellular glucans was inhibited by GIF but not by uncomplexed α-amylase or an unrelated high-molecular-weight glycoprotein. In conclusion, our findings demonstrate that in human saliva, there is a high-molecular-weight glycoprotein-α-amylase complex which is capable of inhibiting GTF and may contribute to control of S. mutans colonization in the oral cavity.

scholarly journals The effect of melanin on the proteolytic potential of blood under alkali esophageal burn

2020 ◽  
Vol 93 (1) ◽  
Author(s):  
Nataliia Chornenka ◽  
Liudmyla Domylivska ◽  
Olga Kravchenko ◽  
Tetiana Koval ◽  
Liza Torgalo ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Proteinase Inhibitors ◽  
Proteolytic Enzymes ◽  
Biological Activities ◽  
Serine Proteinases ◽  
Medical Problems ◽  
Α2 Macroglobulin ◽  
Old Rats ◽  
Hydrolysis Of ◽  
Esophageal Burn

Caustic esophageal burns are among serious medical problems of global significance. Due to a key role in biological processes proteolytic enzymes actively involved in the pathological mechanisms underpinning the development and progression of burn-related complications. Since melanin possesses a broad spectrum of biological activities we have investigated whether this compound can influence the proteolytic activity and level of proteinase inhibitors in the blood of rats with an alkali esophageal burn. Alkaline esophageal burns, which correspond to the first and second degree of burn, were induced by 10% and 20% NaOH, respectively. White, nonlinear, immature (4 weeks old) rats were used in the experiment. Total proteolytic activity was measured using casein as a substrate. The activities of α1- antitrypsin and α2-macroglobulin were measured considering the degree of inhibition of hydrolysis of N-benzoyl-L-arginineethyl ester. The fraction of serine proteinases was obtained by affinity chromatography on a benzamidine sepharose column. The qualitative composition of serine proteinases fraction was analyzed by zymography technique. According to the data obtained, the pathogenesis of alkaline esophageal burn is accompanied by a significant increase in the total proteolytic activity, activity of serine proteinases, and activity of α1-antitrypsin and α2-macroglobulin compared with the control rats. The present results clearly indicated that melanin is able to normalize the proteolytic homeostasis by affecting the activity of serine proteinases and the level of proteinase inhibitors in the plasma of rats with alkali esophageal burns.

scholarly journals Phosphopeptides derived from human salivary acidic proline-rich proteins. Biological activities and concentration in saliva

1990 ◽  
Vol 270 (2) ◽  
pp. 297-304 ◽  
Author(s):  
G Madapallimattam ◽  
A Bennick
Keyword(s):  
Biological Activities ◽  
Human Saliva ◽  
Tooth Surface ◽  
Mineral Formation ◽  
Parotid Saliva ◽  
Whole Saliva ◽  
Tooth Surfaces ◽  
In Vivo Degradation

Human saliva contains a large number of phosphopeptides derived by cleavage of acidic proline-rich proteins (APRPs). These peptides were purified by column chromatography and they constituted 0.5% of APRPs in parotid saliva, but 11% of APRPs in saliva expectorated from the mouth (whole saliva), indicating that there is considerable cleavage of APRPs after secretion from the gland. Similarly to APRP, the phosphopeptides bind Ca2+, but they accounted for only 4% of protein-bound Ca2+ in whole saliva. APRPs as well as the phosphopeptides inhibited formation of hydroxyapatite, but, whereas 19-20 micrograms of APRP was needed for 50% inhibition, only 0.7-3.3 micrograms of purified peptides was needed for the same degree of activity, and the phosphopeptides accounted for 18% of total inhibitory activity in whole saliva. All phosphopeptides adsorbed on hydroxyapatite in vitro, and adsorption of phosphopeptides on tooth surfaces in vivo could also be demonstrated, indicating that they would be able to inhibit unwanted mineral formation on the tooth surface in vivo. Degradation of APRPs after secretion therefore does not lead to a loss of their biological activities.

Expression of Adrenomedullin and its Receptors in Human Salivary Tissue

2004 ◽  
Vol 83 (4) ◽  
pp. 333-337 ◽  
Author(s):  
S. Kapas ◽  
K. Pahal ◽  
A.T. Cruchley ◽  
E. Hagi-Pavli ◽  
J.P. Hinson
Keyword(s):  
Oral Cavity ◽  
Salivary Glands ◽  
Human Saliva ◽  
Oral Epithelium ◽  
Parotid Glands ◽  
Whole Saliva ◽  
Wide Range ◽  
Functional Implications ◽  
High Concentrations

Adrenomedullin is a multifunctional peptide produced by a wide range of different cells and tissues. This study was designed to investigate whether adrenomedullin is present in human saliva and in salivary glands. It was expected that saliva may contain high concentrations of adrenomedullin, which has antimicrobial activity in vitro, which may have functional implications in the oral cavity. Saliva from the submandibular and parotid glands contained higher concentrations of adrenomedullin than did the circulation, but lower concentrations than in whole saliva. This suggests that oral epithelium may contribute the majority of the adrenomedullin peptide found in saliva. Specific adrenomedullin receptors were found in cell lines from the submandibular (HSG) and parotid (HSY) salivary glands. These findings suggest a paracrine/autocrine role for adrenomedullin in these tissues; however, the concentration of adrenomedullin in saliva was insufficient to suggest a significant antimicrobial action in the healthy oral cavity.

Matrix Metalloproteinases (MMP-2 and MMP-9) of the Oral Cavity: Cellular Origin and Relationship to Periodontal Status

1994 ◽  
Vol 73 (8) ◽  
pp. 1397-1406 ◽  
Author(s):  
M. Makela ◽  
T. Salo ◽  
V.-J. Uitto ◽  
H. Larjava
Keyword(s):  
Matrix Metalloproteinases ◽  
Oral Cavity ◽  
Healthy Subjects ◽  
Proteolytic Enzymes ◽  
Host Cells ◽  
Periodontal Treatment ◽  
Primary Role ◽  
Tissue Destruction ◽  
Extracellular Matrix Components ◽  
Whole Saliva

Proteolytic enzymes released by the host cells are associated with the tissue destruction in periodontal diseases. Matrix metalloproteinases (MMPs) have the primary role in this process, since, in concert, they can degrade most of the extracellular matrix components. In the present study, we investigated MMP-2 and MMP-9 in oral fluids of healthy subjects and periodontitis patients and the contributions of different oral cells to the enzyme production. The enzymograms revealed that the main gelatinase in oral rinses, crevicular fluid, and whole saliva migrated at 92 kDa. Activity was also detected at 200 kDa and 130 kDa and minor activity at 86 kDa, 72 kDa, and 40 kDa. Traces of gelatinolytic activity were also detected in pure parotid secretions. The 92-kDa enzyme was identified to MMP-9 and the 200-kDa gelatinase to MMP-2, by means of specific anti-72-kDa antiserum. Gingival keratinocytes produced mainly MMP-9, while gingival and granulation tissue fibroblasts expressed MMP-2. Glandular tissue contained mainly MMP-9, and mRNA for MMP-9 was also found in acinar epithelial cells. Periodontitis patients had significantly higher levels of MMP-9 than healthy subjects. Also, MMP-2 was elevated in periodontitis patients. Periodontal treatment reduced the amount of gelatinases dramatically. This study shows that gelatinases are produced by various cells in the oral cavity. The amount of gelatinases is elevated during periodontal disease, while conventional periodontal treatment efficiently reduces the levels these enzymes. We suggest that MMP-2 and MMP-9 could participate in tissue destruction in periodontitis.

scholarly journals Human Saliva-Mediated Hydrolysis of Eugenyl-β-D-Glucoside and Fluorescein-di-β-D-Glucoside in In Vivo and In Vitro Models

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 172
Author(s):  
Mariusz Dziadas ◽  
Adam Junka ◽  
Henryk Jeleń
Keyword(s):  
Oral Cavity ◽  
Control Sample ◽  
Rapid Test ◽  
Human Saliva ◽  
In Vitro Models ◽  
Microbial Hydrolysis ◽  
Amoxicillin Trihydrate ◽  
Potassium Clavulanate

Eugenyl-β-D-glucopyranoside, also referred to as Citrusin C, is a natural glucoside found among others in cloves, basil and cinnamon plants. Eugenol in a form of free aglycone is used in perfumeries, flavourings, essential oils and in medicinal products. Synthetic Citrusin C was incubated with human saliva in several in vitro models together with substrate-specific enzyme and antibiotics (clindamycin, ciprofloxacin, amoxicillin trihydrate and potassium clavulanate). Citrusin C was detected using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Citrusin C was completely degraded only when incubated with substrate-specific A. niger glucosidase E.C 3.2.1.21 (control sample) and when incubated with human saliva (tested sample). The addition of antibiotics to the above-described experimental setting, stopped Citrusin C degradation, indicating microbiologic origin of hydrolysis observed. Our results demonstrate that Citrusin C is subjected to complete degradation by salivary/oral cavity microorganisms. Extrapolation of our results allows to state that in the human oral cavity, virtually all β-D-glucosides would follow this type of hydrolysis. Additionally, a new method was developed for an in vivo rapid test of glucosidase activity in the human mouth on the tongue using fluorescein-di-β-D-glucoside as substrate. The results presented in this study serve as a proof of concept for the hypothesis that microbial hydrolysis path of β-D-glucosides begins immediately in the human mouth and releases the aglycone directly into the gastrointestinal tract.

scholarly journals Acid Dentin Lysates Increase Amelotin Expression in Oral Epithelial Cells and Gingival Fibroblasts

2021 ◽  
Vol 11 (12) ◽  
pp. 5394
Author(s):  
Jila Nasirzade ◽  
Zahra Kargarpour ◽  
Layla Panahipour ◽  
Reinhard Gruber
Keyword(s):  
Epithelial Cells ◽  
Oral Cavity ◽  
Calcium Binding ◽  
Transforming Growth Factor ◽  
Cell Lineage ◽  
Bone Morphogenetic Protein 2 ◽  
Tooth Surface ◽  
Enamel Matrix Derivative ◽  
Gingival Fibroblasts ◽  
Smad 3

Amelotin (AMTN) is a secretory calcium-binding phosphoprotein controlling the adhesion of epithelial cells to the tooth surface, forming a protective seal against the oral cavity. It can be proposed that signals released upon dentinolysis increase AMTN expression in periodontal cells, thereby helping to preserve the protective seal. Support for this assumption comes from our RNA sequencing approach showing that gingival fibroblasts exposed to acid dentin lysates (ADL) greatly increased AMTN expression. In the present study, we confirm that acid dentin lysates significantly increase AMTN in gingival fibroblasts and extend this observation towards the epithelial cell lineage by use of the HSC2 oral squamous and TR146 buccal carcinoma cell lines. AMTN immunostaining revealed an intensive signal in the nucleus of HSC2 cells exposed to acid dentin lysates. Acid dentin lysates mediate their effect via the transforming growth factor (TGF)-β type 1 receptor kinase as the antagonist SB431542 abolished the expression of AMTN in the epithelial cells and fibroblasts. Similar to what is known for fibroblasts, acid dentin lysate increased Smad-3 phosphorylation in HSC2 cells. HSC2 cells also respond to the AMTN-stimulating activity of the dentin lysate when adsorbed to gelatin. When simulating regenerative approaches, enamel matrix derivative, TGF-β1, and bone morphogenetic protein-2 also caused a robust increase in SB431542-dependent AMTN expression in HSC2. Taken together, we show here that acid dentin lysate uses the TGF-β-depended signaling pathway to support the AMTN expression in epithelial cells, possibly helping in maintaining the protective seal against the oral cavity.

scholarly journals Non-conventional affinity chromatography of serine proteinases and their inhibitors.

2003 ◽  
Vol 50 (3) ◽  
pp. 765-773 ◽  
Author(s):  
Antoni Polanowski ◽  
Anna Wilimowska-Pelc ◽  
Jolanta Kowalska ◽  
Joanna Grybel ◽  
Monika Zelazko ◽  
...  
Keyword(s):  
Affinity Chromatography ◽  
Proteinase Inhibitor ◽  
Proteolytic Enzymes ◽  
Trypsin Inhibitors ◽  
General Description ◽  
Separation Procedure ◽  
Serine Proteinases ◽  
High Concentration ◽  
Porcine Pancreas ◽  
Nacl Concentration

From among a wide variety of protein purification techniques affinity chromatography has proved to be particularly effective for separation of proteolytic enzymes and their inhibitors. In this article, following a general description of affinity adsorbents used for purification of proteinases, we overview a simple separation procedure for some serine proteinases and their inhibitors by way of affinity chromatography in the presence of high NaCl concentration. It has been shown that some highly specific trypsin inhibitors exhibit also antichymotrypsin activity when high concentration of Na(+) but not K(+) or Li(+) ions are present in the reaction mixture. Taking advantage of this phenomenon the virgin forms of trypsin inhibitors from squash seeds, Kazal-type inhibitor from porcine pancreas and alpha(1)-proteinase inhibitor from human and sheep plasma, as an example, were separated using immobilized chymotrypsin or its inactive derivative methylchymotrypsin in the presence of 5 M NaCl.

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