Two-dimensional electrophoresis of human parotid salivary proteins from normal and connective tissue disorder subjects using immobilised pH gradients.
Two-dimensional electrophoretic analysis of human salivary proteins using immobilised pH gradients in the first dimension, thin-layer gradient horizontal sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second, and modified staining procedures has resulted in a substantial improvement in their resolution. Unlike carrier ampholyte-based techniques, immobilised pH gradients prevent the loss of proteins of pI greater than 8; accordingly, basic components, including basic proline-rich proteins, can now be resolved. A two-dimensional map showing the locations and identities of most of the major proteins has been constructed. Narrow-range pH gradients can be constructed to give increased resolution of proteins of particular interest. By means of a pH 3.5-5.0 gradient, the abnormal salivary proteins associated with connective tissue disorders were found to be a highly heterogeneous group of pI approximately 3.75-4.75 and Mr approximately 32,000; although low levels occurred in some normal individuals, there was less heterogeneity (pI approximately 3.75-4.25). The technique should form a base for future structural, functional, and clinical studies on human salivary proteins.
Beeley
J A
JA
Dental School Oral Biochemistry Unit, University of Glasgow, UK.
Khoo
K S
KS
Lamey
P J
PJ
D001172:Arthritis, Rheumatoid / Q000503:physiopathology; D003240:Connective Tissue Diseases / Q000503:physiopathology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods; D008970:Molecular Weight; D010306:Parotid Gland / Q000378:metabolism*; D012016:Reference Values; D012471:Salivary Proteins and Peptides / Q000302:isolation & purification* / Q000378:metabolism; D012859:Sjogren's Syndrome / Q000503:physiopathology; D013194:Staining and Labeling
D012471:Salivary Proteins and Peptides