Camphor Seeds Ribosome Inactivating Protein Stability Study

Ribosome inactivating protein is a protein of RNA N-glycosylase activity, which widely exists inhigher plants root, leaf and seed cells and has excellent performance in anti-tumor, anti-HIV, and animaland plant virus resistance. Cinnamomum camphora seeds contain a lot of ribosome inactivating protein,and Cinnamomum camphora trees are widely distributed in our country. But at present we don’t use theCinnamomum camphora well which is not only a waste of the resource material, but also a kind ofenvironment pollution.This paper use the Cinnamomum camphora seed as the research object, with the method of using theacetone and ammonium sulfate tor protein precipitation, ligand affinity chromatography for separation andpurification of ribosome inactivating protein in the Cinnamomum camphora seed. After interacting with thehepatocarcinoma cells, we will study the properties of the Cinnamomum camphora seed. We study byprocessing it with MgCl2and protease inhibitors, and then analyzing its properties with SDS-PAGEelectrophoresis, in order to understand the properties of the ribosome inactivating protein, thetransformation relation and condition between two kinds of ribosome inactivating protein in Cinnamomumcamphora seeds. The main results are as follows:1.Identified a kind of extraction technology which can rapidly and conveniently extract Cinnamomumcamphora seeds ribosome inactivating protein. Through the electrophoresis identification and interactionwith cancer cells, we are sure that the extracted protein is the ribosome inactivating protein2.Remove grease and other impurities in the extraction process. Using the acetone precipitation ofprotein concentration enrichment is much better than ammonium sulfate precipitation and uses less time.But the disadvantages are that the inactivation of protein is more visible. Therefore, in terms of purifying the ribosome inactivating protein we can use the ammonium sulfate precipitation to concentrate, but bydifferent conditions of processing we can use the acetone precipitation.3.Compared with the degradation trend of the ribosome inactivating protein in the Cinnamomumcamphora under the natural condition, the degradation trend after treating by the MaCl2and EDTA–PMSF is significant different. The dump toxic protein degradation in camphorin in the crude extractionwhich is treated with MaCl2is greater than that of no reagent treated crude extract. But with EDTA-PMSFprocessing, because the protein is protected, the camphorin which is generated is much greater tan that ofno reagent treated crude extract.4.The transformation relation between the Cinnamomin and Camphorin is proportional to the externalconditions, such as time and temperature. It has certain consistency with the fruit cell ageing process, whichcan be used to illustrate the transformation relationship between fruit cell aging process.