HIGH MOLECULAR WEIGHT SUBSTANCES WHICH INHIBIT CALCIUM OXALATE CRYSTAL GROWTH
2: Isolation and Identification of a Calcium Oxalate Crystal Growth Inhibitor from Rat Liver
1978 Volume 69 Issue 10 Pages 1342-1348
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1978 Volume 69 Issue 10 Pages 1342-1348
A calcium oxalate crystal growth inhibitory substance was obtained by 3M MgCl2 extraction and ethanol precipitation. The density of this substance was found to be very high by applying the material to CsCI buoyant density gradient centrifugation.
The rat liver was homogenized in water and heated at 80°C for 30min. This active substance was applied to a DEAE-cellulose column and eluted from EADE by increasing the concentration of NaCl in the buffer. Each fraction was assayed for inhibitory activity, protein, RNA, DNA, and uronic acid.
The active fraction from DEAE was further purified by preparative disc gel electrophoresis using 3% polyacrylamide 0.5% agarose composite gel. The substance was electrophoresed faster than bromophenol blue. It became clear since this substance is very acidic.
These two characteristics along with the elution pattern from DEAF-cellulose column suggested that this material is RNA. This hypothesis was confirmed by the ultraviolet absorption spectra and the digestion experiment with RNases.
