Large-scale purification and characterization of the major phosphoproteins and mucins of human submandibular-sublingual saliva.
The major components of human submandibular-sublingual saliva (HSMSL) are mucins, amylases, cystatins, proline-rich proteins and statherin. Structure-function studies of these molecules have been hampered by the small amounts of purified materials that can be isolated from human secretions. The present study describes an integrated purification protocol for the large-scale preparation of many of these molecules. To dissociate partially heterotypic complexes among salivary molecules, HSMSL was initially fractionated into four pools by gel filtration with 6 M-guanidine hydrochloride. Subsequent fractionation of these four pools by gel-filtration and ion-exchange chromatography resulted in the purification of high- and low-Mr mucins, neutral and acidic cystatins, acidic and basic proline-rich proteins and statherin. Many variants or isoforms of these salivary molecules have been identified and biochemically characterized. Biochemical studies indicated that the low-Mr mucin exists as two isoforms which vary in their sialic acid to fucose ratios. Three isoforms of acidic cystatin S were characterized which differ in their phosphate content. Two isoforms of a basic proline-rich peptide were identified; the smaller peptide was a truncated form missing the first seven amino acids.
Ramasubbu
N
N
Department of Oral Biology, School of Dental Medicine, State University of New York, Buffalo 14214.
Reddy
M S
MS
Bergey
E J
EJ
Haraszthy
G G
GG
Soni
S D
SD
Levine
M J
MJ
D000328:Adult; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D015891:Cystatins / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry / Q000302:isolation & purification*; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification*; D011480:Protease Inhibitors / Q000378:metabolism; D012463:Saliva / Q000737:chemistry*; D013361:Sublingual Gland / Q000737:chemistry*; D013363:Submandibular Gland / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000037:antagonists & inhibitors
D000596:Amino Acids; D015891:Cystatins; D009077:Mucins; D010750:Phosphoproteins; D011480:Protease Inhibitors; D013438:Sulfhydryl Compounds