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Association of endogenous substrate with solubilized bovine brain sialidase
Author(s) -
Schengrund C.L.,
Repman M. A.
Publication year - 1986
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490150207
Subject(s) - sialidase , endogeny , ganglioside , substrate (aquarium) , enzyme , sialic acid , chemistry , biochemistry , protease , membrane , solubilization , neuraminidase , biology , ecology
Nonidet P4O solubilized up to 90% of the sialidase, active towards added ganglioside substrate, that was associated with the total membrane fraction prepared from gray matter of bovine brains. Solubilized sialidase acted upon endogenous substrate (sialic acid containing compounds solubilized with the enzyme), hydrolyzing approximately 50% of the readily available sialosyl residues within 20 min. During a 2‐hr reaction time 80% of the polysialylated gangliosides solubilized with the enzyme were acted upon. A 20‐min lag was observed before sialidase acted upon added ganglioside substrate. The lag could be reduced to less than 2 min when the enzyme was allowed to act on endogenous substrate prior to exposure to exogenous substrate, suggesting that the solubilized enzyme acted preferentially on endogenous substrate. A protease inhibitor prevented much of the 86% loss of activity towards added substrate that was seen when the enzyme was stored at 4°C for 6 days; activity towards endogenous substrate decreased only 34%.