Inhibition of calcium phosphate precipitation by human salivary acidic proline-rich proteins: structure-activity relationships.
Absence of precipitation of calcium phosphate salts onto tooth surfaces from human saliva, which is supersaturated with respect to calcium phosphate salts, has been attributed in part to the presence in the salivary secretions of a group of acidic proline-rich phosphoproteins (PRP). These macromolecules are considered to act by adsorbing onto dental enamel where they inhibit surface-induced precipitation of calcium phosphate salts. The inhibitory activity is known to be associated primarily with the amino-terminal region of the PRP. The aim of this study was to determine the features of the primary structure of this molecular segment responsible for inhibitory activity. The 30-residue, amino-terminal segment of PRP-3, which contains the two phosphoserines and 11 of the 13 carboxyl groups present in PRP-3, was obtained by tryptic digestion. This peptide, designated PRP-3(TI), was treated with thermolysin to give the monophosphopeptides, Val-PSer-Gln-Glu-Asp-Val-Pro and Leu-Val-Ile-Ser-Asp-Gly-Gly-Asp-PSer-Glu-Gln, and with alkaline phosphatase to give the dephosphorylated analog, PRP-3(TI)DP. The inhibitory activities of PRP-3(TI) and the derived peptides, a synthetic peptide, phosphoseryl-phosphoserine (PSer-PSer), and O-phosphoserine (PSer), were determined using an assay based on inhibition of seeded precipitation of calcium phosphate. Inhibitory activities, expressed as concentrations of inhibitors required to give standard inhibitory activities, were PRP-3(TI), 0.59 microM; PSer-PSer, 3.5 microM; the two monophosphopeptides, 29 and 32.5 microM; PRP-3(TI)DP, 56 microM; PSer, 329 microM.(ABSTRACT TRUNCATED AT 250 WORDS)
D002130:Calcium Phosphates / Q000378:metabolism*; D011232:Chemical Precipitation; D003743:Dental Enamel / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D010446:Peptide Fragments / Q000302:isolation & purification; D010455:Peptides / Q000378:metabolism*; D055232:Proline-Rich Protein Domains; D012463:Saliva / Q000378:metabolism*; D055273:Salivary Proline-Rich Proteins; D013329:Structure-Activity Relationship; D013820:Thermolysin
D002130:Calcium Phosphates; C524536:PRH1 protein, human; D010446:Peptide Fragments; D010455:Peptides; D055273:Salivary Proline-Rich Proteins; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; D013820:Thermolysin; C485829:calcium phosphate, dibasic, anhydrous