Method for total protein fractionation and analysis of human saliva
AssigneeThe United States of America as represented by the Department of Health, Education and Welfare
InventorRobert I. Henkin
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Abstract
A method for the total protein fractionation of saliva and for isolating a novel zinc protein found to be a component thereof is disclosed. Saliva was fractionated using three protein parameters and zinc concentration by a molecular sieving process. The zinc protein was isolated and purified from the saliva of subjects with normal taste acuity by gel filtration and ion exchange chromatography maximizing these parameters. The zinc protein has a molecular weight of 37,000 and does not appear to have subunits. It is composed of 8% histidine residues and has two moles of zinc per mole of protein. A decreased concentration of the protein in saliva has been associated with abnormal taste acuity.
Claims37
A method for analyzing saliva to determine the total protein content thereof comprising the steps of:
- fractionating the protein components of said saliva on a molecular sieve, said protein components separating as a function of their molecular weights and aromaticity;
- collecting separate aliquots of said fractionated saliva;
- measuring protein parameters dependent upon (1) the amount of the peptide chromophore;
- (2) the amount of tyrosine and tryptophan;
- and (3) the quantum yield of tryptophan;
- and (4) the zinc concentration in each aliquot;
- recording said measured parameters to provide a total protein fractionation record.
A method for isolating and identifying a zinc protein from human saliva comprising the steps of:
- fractionating the protein components of said saliva on a molecular sieve, said protein components separating as a function of their molecular weights and aromaticity;
- collecting separate aliquots of said fractionated saliva;
- measuring protein parameters dependent upon (1) the amount of the peptide chromophore;
- (2) the amount of tyrosine and tryptophan;
- and (3) the quantum yield of tryptophan and (4) the zinc concentration in each aliquot;
- selecting those aliquots wherein the zinc concentration to protein parameters ratios are maximum;
- subjecting the selected aliquot fraction to a combined molecular sieve and ion exchange component separation;
- collecting separate aliquots of said separated material;
- measuring protein parameters dependent upon (1) the amount of the peptide chromophore;
- (2) the amount of tyrosine and tryptophan;
- and (3) the quantum yield of tryptophan and (4) the zinc concentration in each aliquot;
- selecting those aliquots wherein the zinc concentration to protein parameters are maximum;
- subjecting the selected aliquot fraction to a final purification by ion exchange;
- collecting separate aliquots of said material;
- measuring the protein parameters dependent upon (1) the amount of the peptide chromophore;
- (2) the amount of tyrosine and tryptophan;
- and (3) the quantum yield of tryptophan and (4) the zinc concentration in each aliquot;
- selecting those aliquots wherein the zinc concentration to protein parameter ratios are constant and the ratios of the parameters are constant to each other.
A method for diagnosing zinc deficiency and evaluating zinc nutriture comprising the steps of:
- collecting saliva and subjecting said saliva to a total protein fractionation while measuring protein parameters dependent upon (1) the amount of the peptide chromophore;
- (2) the amount of tyrosine and tryptophan;
- and (3) the quantum yield of tryptophan;
- and (4) the zinc concentration;
- recording said measured results for evaluation of said recorded results against normal values.
A method for diagnosing diseases of protein synthesis or diseases characterized by the presence of abnormal proteins or abnormal protein synthesis comprising the steps of:
- collecting saliva and subjecting said saliva to a total protein fractionation while measuring protein parameters dependent upon (1) the amount of the peptide chromophore;
- (2) the amount of tyrosine and tryptophan;
- and (3) the quantum yield of tryptophan;
- and (4) the zinc concentration;
- recording said measured results for evaluation of said recorded results against normal values.
A method for diagnosing taste dysfunction comprising the steps of:
- collecting saliva and subjecting said saliva to a total protein fractionation while measuring protein parameters dependent upon (1) the amount of the peptide chromphore;
- (2) the amount of tyrosine and tryptophan;
- and (3) the quantum yield of tryptophan;
- and (4) the zinc concentration;
- recording said measured results for evaluation of said recorded results against normal values.
Dependencies are inferred from the claim text.