Electrophoretic analysis of stimulated cat parotid saliva

The proteins of parasympathetically stimulated cat parotid saliva were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and high resolution two-dimensional electrophoresis (2-DE). SDS-PAGE revealed up to 30 polypeptide bands in microliter volumes of unconcentrated saliva. The patterns were highly reproducible and characterized by prominent bands of M(r) 57,000, M(r) 30,000 and M(r) 15,000. The major protein (M(r) 30,000) appeared as a dimer (M(r) 60,000) when electrophoresed under non-reducing conditions but dissociated into its monomeric form when the SDS concentration of the denatured samples was increased from 1 to 5%. This indicates a noncovalent association. The protein patterns of saliva from different cats differed slightly but sequential samples from the same cat (collected during 90 min of stimulation) showed little change in protein pattern apart from a fall in total protein content. Following 2-DE, the major protein (M(r) 30,000) appeared as a complex array of at least eight spots in two tiers (pI 5.2-6.2; M(r) 28,000 and 32,000). The characteristics of this protein are discussed with reference to allergy to cats.