Trafficking and Postsecretory Events Responsible for the Formation of Secreted Human Salivary Peptides

Messana, Irene; Cabras, Tiziana; Pisano, Elisabetta; Sanna, Maria Teresa; Olianas, Alessandra; Manconi, Barbara; Pellegrini, Maria Grazia; Paludetti, Gaetano; Scarano, Emanuele; Fiorita, Antonella; Agostino, Stefania; Contucci, Alessia Maria; Calò, Lea; Picciotti, Pasqualina Maria; Manni, Antonio; Bennick, Anders; Vitali, Alberto; Fanali, Chiara; Inzitari, Rosanna; Castagnola, Massimo

doi:10.1074/mcp.m700501-mcp200

Cited by 111 publications

(201 citation statements)

References 49 publications

Supporting

Mentioning

184

Contrasting

Order By: Relevance

“…However, contribution of the submandibular/sublingual glands was slightly higher than parotid glands because saliva was collected from the floor of the mouth close to orifices of Wharton's ducts. HPLC-ESI-MS analysis of whole human saliva allows detecting more than 120 salivary components (25). In the present study, we investigated the following already characterized salivary proteins: salivary acidic proline-rich phosphoproteins, histatins, ␣-defensins, salivary cystatins (Table I), statherin, proline-rich peptide P-B, and beta-thymosins (Table II), as well as several derivatives generated by post-translational modifications occurring before and after secretion, such as phosphorylated isoforms and cleavage products.…”

Section: Resultsmentioning

confidence: 99%

Alterations of the Salivary Secretory Peptidome Profile in Children Affected by Type 1 Diabetes

Cabras

Pisano

Mastinu

et al. 2010

Molecular & Cellular Proteomics

Self Cite

View full text Add to dashboard Cite

The acidic soluble fraction of whole saliva of type 1 diabetic children was analyzed by reversed phase (RP) 1 -HPLC-ESI-MS and compared with that of sex-and age-matched control subjects. Salivary acidic proline-rich phosphoproteins (aPRP), histatins, ␣-defensins, salivary cystatins, statherin, proline-rich peptide P-B (P-B), betathymosins, S100A8 and S100A9*(S100A9* corresponds to S100A9 vairant lacking the first four amino acids), as well some naturally occurring peptides derived from salivary acidic proline-rich phosphoproteins, histatins, statherin, and P-B peptide, were detected and quantified on the basis of the extracted ion current peak area. The level of phosphorylation of salivary acidic proline-rich phosphoproteins, histatin-1 (Hst-1), statherin and S100A9* and the percentage of truncated forms of salivary acidic proline-rich phosphoproteins was also determined in the two groups. The study revealed that statherin, proline-rich peptide P-B, P-C peptide, and histatins, were significantly less concentrated in saliva of diabetic subjects than in controls, while concentration of ␣-defensins 1, 2 and 4 and S100A9* was higher. The low concentration of P-C peptide was paralleled by high levels of some of its fragments. On the whole, the study highlighted the severe impairment of the repertoire of peptides involved in the safeguard of the oral cavity in children who have diabetes, as well as an higher concentration of the proinflammatory mediator S100A9* with respect to healthy children. Molecular & Cellular Proteomics 9:2099 -2108, 2010.Type 1 diabetes mellitus is a disease with universal distribution, affecting all populations (1) with an incidence increasing in Europe (2). Because type 1 diabetes involves many organs and tissues, signs and symptoms of diabetes can occur in the oral cavity. Indeed, several studies have shown that the prevalence, severity, and progression of periodontal diseases are significantly increased in diabetics, and the pathology is considered an important risk factor for periodontitis (3). The study of Lalla et al. (4) showed an association between diabetes and the increased risk for periodontal destruction even very early in life. Flow rate and composition of saliva are crucial for the maintenance of oral cavity health, and both have been found altered in diabetic subjects, although with contradictory findings. For instance, several studies reported a reduced resting salivary flow rate in adults and children who have type 1 diabetes with respect to healthy subjects (5-10), but it was also observed that stimulated saliva of diabetics and controls did not significantly differ (11,12). Total protein concentration of saliva was found either increased or not affected by the diabetic status. Lopez et al. (9) reported higher urea and total protein salivary content in diabetic children than in controls. Conversely, a study on 35 adult insulin-dependent diabetics demonstrated that they did not show a decreased saliva concentration of several innate antimicrobial factors (lysozyme, lactoferri...

show abstract

Section: Resultsmentioning

confidence: 99%

Alterations of the Salivary Secretory Peptidome Profile in Children Affected by Type 1 Diabetes

Cabras

Pisano

Mastinu

et al. 2010

Molecular & Cellular Proteomics

Self Cite

View full text Add to dashboard Cite

show abstract

“…Proteolytic Enzymes in Whole Saliva-It is of interest that there are a number of proteases associated with human salivary glands that are responsible for post-translational processing of some salivary proteins (43,44). One of the protein families partially fragmented in the gland are the basic PRPs.…”

Section: Discussionmentioning

confidence: 99%

“…This fragment corresponds to the dephosphorylated, doubly charged parent ion having a calculated m/z value of (1240.89 Ϫ 98.0 ϩ 2.0)/2 ϭ 572. 44. Such dephosphorylated parent ions in the MS/MS spectrum are typical and characteristic for phosphorylated peptides and help in their identification (35,36).…”

Section: Extent Of Proteolysis In the Oralmentioning

confidence: 99%

Identification of Lys-Pro-Gln as a Novel Cleavage Site Specificity of Saliva-associated Proteases

Helmerhorst

Sun

Salih

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

The nonsterile environment of the oral cavity facilitates substantial proteolytic processing, not only of resident salivary proteins but also of dietary proteins. To gain insight into whole saliva enzymatic processes, the in vivo generated peptides in this oral fluid were subjected to nano-flow liquid chromatography electrospray ionization tandem mass spectrometry. The 182 peptides identified were predominantly derived from acidic and basic proline-rich proteins, statherin, and histatins. The proteolytic cleavages in the basic proline-rich proteins occurred preferentially after a Gln residue with predominant specificity for the tripeptide Xaa-Pro-Gln, where Xaa in the P 3 position was mostly represented by Lys. Using the synthetic substrates LysPro-Gln-pNA and Gly-Gly-Gln-pNA, the overall K m values were determined to be 97 ؎ 7.7 and 611 ؎ 28 M, respectively, confirming glutamine endoprotease activity in whole saliva and the influence of the amino acids in positions P 2 and P 3 on protease recognition. The pH optimum of Lys-Pro-Gln-pNA hydrolysis was 7.0, and the activity was most effectively inhibited by antipain and 4-(2-aminoethyl) benzenesulfonyl fluoride, was metal ion-dependent, and not inhibited by cysteine protease inhibitors. A systematic evaluation of enzyme activities in various exocrine and nonexocrine contributors to whole saliva revealed that the glutamine endoprotease is derived from dental plaque and likely microbial in origin. The P 1 site being occupied by a Gln residue is a nonarchetype with respect to known proteases and indicates the presence of novel glutamine-specific endoprotease(s) in oral fluid.

show abstract

“…This relationship between proteins and their hydrolytic products has fueled the emergence of peptidomics as a subdiscipline of proteomics. Human biofluids such as blood (3), cerebrospinal fluid (4), saliva (5,6), tears (7), and urine (8) have been analyzed for endogenous peptides. As naturally occurring peptides reflect both the protein content of a tissue and a specific configuration of the proteolytic machinery, they represent a promising target for biomarker discovery (9 -13).…”

mentioning

confidence: 99%

Mechanistic Peptidomics: Factors That Dictate Specificity in the Formation of Endogenous Peptides in Human Milk

Guerrero

Dallas

Contreras

et al. 2014

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

An extensive mass spectrometry analysis of the human milk peptidome has revealed almost 700 endogenous peptides from 30 different proteins. Two in-house computational tools were created and used to visualize and interpret the data through both alignment of the peptide quasi-molecular ion intensities and estimation of the differential enzyme participation. These results reveal that the endogenous proteolytic activity in the mammary gland is remarkably specific and well conserved. Certain proteins-not necessarily the most abundant ones-are digested by the proteases present in milk, yielding endogenous peptides from selected regions. Our results strongly suggest that factors such as the presence of specific proteases, the position and concentration of cleavage sites, and, more important, the intrinsic disorder of segments of the protein drive this proteolytic specificity in the mammary gland. As a consequence of this selective hydrolysis, proteins that typically need to be cleaved at specific positions in order to exert their activity are properly digested, and bioactive peptides encoded in certain protein sequences are released. Proteins that must remain intact in order to maintain their activity in the mammary gland or in the neonatal gastrointestinal tract are unaffected by the hydrolytic environment present in milk. These results provide insight into the intrinsic structural mechanisms that facilitate the selectivity of the endogenous milk protease activity and might be useful to those studying the peptidomes of other biofluids. Molecular &

show abstract

Trafficking and Postsecretory Events Responsible for the Formation of Secreted Human Salivary Peptides

Cited by 111 publications

References 49 publications

Alterations of the Salivary Secretory Peptidome Profile in Children Affected by Type 1 Diabetes

Alterations of the Salivary Secretory Peptidome Profile in Children Affected by Type 1 Diabetes

Identification of Lys-Pro-Gln as a Novel Cleavage Site Specificity of Saliva-associated Proteases

Mechanistic Peptidomics: Factors That Dictate Specificity in the Formation of Endogenous Peptides in Human Milk

Contact Info

Product

Resources

About