A quantitative isolation method for gangliosides from small sizes of samples has been developed. Total lipids were separated into gangliosides and other lipids using Phenyl Sepharose column chromatography. Gangliosides were recovered in a yield of \(\mathbf{9 9 \%}\). Determination of gangliosides w
Characterization of sialidase activity in mouse synaptic plasma membranes and its age-related changes
โ Scribed by Dr. M. Saito; Y. Tanaka; C.-P. Tang; R. K. Yu; S. Ando
- Publisher
- John Wiley and Sons
- Year
- 1995
- Tongue
- English
- Weight
- 585 KB
- Volume
- 40
- Category
- Article
- ISSN
- 0360-4012
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โฆ Synopsis
Sialidase activity in synaptic plasma membranes (SPM) isolated from C57BL/6 mouse brain was examined using exogenous ganglioside substrates. The enzyme activity directed toward GM3 showed sharp pH dependency with optimal pH of 4.0, and was greatly enhanced by Triton CF-54, Nonidet P-40 or CHAPS. The apparent K, and V,,, values for enzyme activity in SPM were 11 pM and 164 pmollmg protein/min, respectively. Examination of sialidase activities in subcellular fractions of brain tissues showed the enrichment of enzyme activity in SPM prepared from either young adult or senescent mice. Substrate specificity of SPM sialidase was compared with that of myelin sialidase using delipidated, solubilized enzyme preparations. The SPM sialidase hydrolyzed GDla more effectively as compared with the myelin enzyme. While SPM sialidase could hydrolyze GMl, the hydrolytic rate by the SPM enzyme was significantly lower than that by the myelin enzyme. The sialidase activity in SPM decreased with increasing age; activity was highest between the ages of 4-7 months, decreased to a relatively constant level between 13-25 months, and reached its lowest level at 31 months. These results demonstrate that SPM contain a distinct sialidase activity which is regulated in an age-dependent manner.
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