Anti-peptide antibodies reactive with epitopic domains of porcine amelogenins at the C-terminus.
This was an immunological investigation of the processing of porcine amelogenins in situ. Rabbit and rat anti-peptide sera reacted specifically with the hydrophilic segment of the intact amelogenins at the C-terminus. The immunogens used were the synthetic peptides: (a) C13 composed of PATDKTKREEVDC and (b) C25 composed of MQSLLPDLPLEAWPATDKTKREEVD. These peptides correspond to the C-terminal 12- and 25-residue segments of porcine amelogenin, respectively. Cystine was introduced at the C-terminus of C12 for KLH-binding (C13). Western blot analysis disclosed that: (i) both rabbit and rat anti-C13 sera reacted selectively with the 25-kDa porcine amelogenin and three other minor components (27, 22 and 18 kDa); (ii) anti-C25 peptide sera, additionally, reacted with the 23-kDa amelogenins (a degradation derivative of the 25-kDa protein, lacking the 12-residue segment at the C-terminus) and as trace components, 20-, 16- and 14-kDa moieties. Importantly, all the proteins reactive with the anti-C13 serum were concentrated in the outer secretory enamel adjacent to the ameloblasts, decreasing significantly in the underlying inner secretory enamel. Immunohistochemical studies applying the anti-peptide sera to the developing tooth germs of a minipig also confirmed the localization of reactivity in the outer secretory region. Neither anti-peptide serum reacted with porcine non-amelogenins, serum proteins nor dentine matrix proteins at the dilutions tested. however, it was found that both the anti-C13 and C25 sera reacted with human keratin.(ABSTRACT TRUNCATED AT 250 WORDS)
Aoba
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Forsyth Dental Center, Boston, MA 02115.
Shimoda
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Akita
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Holmberg
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Taubman
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D053523:Amelogenin; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001224:Aspartic Acid / Q000276:immunology; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003743:Dental Enamel / Q000737:chemistry / Q000276:immunology; D003746:Dental Enamel Proteins / Q000276:immunology* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D008715:Methionine / Q000276:immunology; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology* / Q000302:isolation & purification*; D011392:Proline / Q000276:immunology; D012967:Sodium Dodecyl Sulfate; D013552:Swine; D014083:Tooth Germ / Q000737:chemistry* / Q000276:immunology*
D053523:Amelogenin; C507201:Amelx protein, rat; D000596:Amino Acids; D000906:Antibodies; D003746:Dental Enamel Proteins; D000939:Epitopes; D010446:Peptide Fragments; D001224:Aspartic Acid; D012967:Sodium Dodecyl Sulfate; D011392:Proline; D008715:Methionine