Identification of zinc proteins in rat parotid saliva.
The presence of unique zinc-binding proteins in human saliva is well documented. These observations have not, however, been extended to other species. The rat has been used extensively to study the salivary gland and its secretion, and it is therefore important to determine if the spectrum of zinc-binding proteins in this experimental model resembles that found in humans. To begin the analysis of zinc-binding proteins in stimulated rat parotid saliva, the saliva was fractionated by DEAE Sephadex and Sepharose 6B chaelate chromatography and the protein patterns analysed by electrophoresis. Zinc-binding proteins from the parotid saliva were identified by incubating Western blots with 65Zn and identifying any bound zinc by autoradiography. Comparison of the autoradiograms with the Coomassie blue-stained filter revealed several proteins with zinc-binding capacity. Isolation of the major zinc-binding proteins revealed an amino acid composition of proline 28%, glutamine 19% and glycine 15%, which is consistent with the amino acid composition of rat salivary acidic proline-rich protein. In addition to the proline-rich proteins, one other zinc-binding protein was analysed. The N-terminal sequence of this protein was found to bear a striking similarity (16 out of 20 amino acids) to secreted carbonic anhydrase VI of the mouse, a known zinc-binding protein. These data demonstrate that rat acidic proline-rich proteins, having an amino acid composition similar to that in humans, have zinc-binding potential. The data also confirm previous reports suggesting secreted carbonic anhydrase in rat parotid saliva.
Etzel
K R
KR
Department of Microbiology/Biochemistry, University of Pittsburgh School of Dental Medicine 15261-1964. USA.
Hempel
J D
JD
Koepsel
R R
RR
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002256:Carbonic Anhydrases / Q000032:analysis; D002352:Carrier Proteins / Q000032:analysis*; D002417:Cattle; D002845:Chromatography / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008297:Male; D008667:Metalloproteins / Q000032:analysis*; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D012471:Salivary Proteins and Peptides / Q000032:analysis / Q000737:chemistry*; D015032:Zinc / Q000032:analysis*
D000596:Amino Acids; D002352:Carrier Proteins; D008667:Metalloproteins; D012471:Salivary Proteins and Peptides; C017800:zinc-binding protein; D002256:Carbonic Anhydrases; C477051:carbonic anhydrase VI; D015032:Zinc