Sialidase in rat salivary glands

In this study, we elucidated some characteristics of soluble sialidases in the rat salivary glands, brain, liver and kidney. 1) Soluble sialidases in the submandibular and parotid glands were inactivated by about 75 and 60%, respectively, and the enzymes in the brain, liver and kidney were also inactivated by 16-48%, when preincubated at 40•Ž for 1h. When preincubated at 60•Ž, the enzymes in all tissues were completely inactivated within 10 min. 2) Ca2+ ion in low concentrations tended to activate soluble sialidases in both the submandibular and parotid glands, but had no apparent effect on the enzymes in the brain, liver and kidney. The enzyme in the submandibular gland tended to be activated also by low concentrations of Mg2+. Both Cu2+ and Hg2+ ions caused a marked inhibition on the soluble sialidases in all tissues. 3) The molecular weight of soluble sialidases in the submandibular and parotid glands was determined to be about 68,000 and 46,000, respectively, by means of Sephadex G-200 gel filtration. The molecular weight of the enzymes in the brain and liver was almost similar to that of the enzyme in the parotid gland. In the kidney, soluble sialidase was separated into two enzymes, one having high (more than 500,000) and the other having low (about 46,000) molecular weight. 4) By isoelectric focusing analysis, three isozymes were detected in the submandibular gland and brain sialidases and two isozymes detected in the enzymes of other tissues. The isoelectric point of the major isozymes in the submandibular and parotid glands was 6.4 and 6.9 respectively. In the brain, liver and kidney sialidases, the isozymes with isoelectric point in the range of 4.4 to 6.7 were detected.