Isolation and characterization of six proteins from rabbit parotid saliva belonging to a unique family of proline-rich proteins.
Proline-rich proteins are major components of salivary secretion from humans non-human primates, rats, hamsters and rabbits. They are also synthesized in mice in response to chronic stimulation by beta agonists. This study to provide an understanding of the structural and genetic relationships within these families of proteins to determine the possible function of the proline-rich proteins. Rabbit parotid saliva was collected and proline-rich proteins were affinity purified using goat antibodies to human proline-rich proteins. Purification was achieved by repeated cation exchange chromatography on a Mono S column a Fast Protein Liquid Chromatography system. Six basic proline-rich proteins were purified. The apparent molecular weights were between 75,000 and 125,000, based on their mobilities in sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Glycine, glutamine (and glutamate) and proline accounted for 79-87% of total amino acids in all proteins, but proline was present in smaller amounts (17-21%) than in proline-rich proteins from other species. All proteins were glycosylated but not phosphorylated. Circular dichroism of two proline-rich proteins, MS7A and MS5B, indicated the absence of secondary structure. The N-terminal sequences of three proteins electro-eluted after preparative gel electrophoresis were determined. A high degree of similarity was found in various regions of mouse, rat, monkey and human proline-rich proteins. Rabbits thus synthesize constitutively a family of proteins that are immununologically and structurally related to proline-rich proteins other species.
Department of Biochemistry, University of Toronto, Ontario, Canada.
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Spielman
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Bennick
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015153:Blotting, Western; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D010306:Parotid Gland / Q000032:analysis / Q000378:metabolism*; D010455:Peptides / Q000302:isolation & purification*; D055232:Proline-Rich Protein Domains; D011817:Rabbits; D012463:Saliva / Q000032:analysis*
D000596:Amino Acids; D010455:Peptides