Large-scale preparation of two new ribosome-inactivating proteins--cinnamomin and camphorin from the seeds of Cinnamomum camphora.
An improved method for large-scale preparation of cinnamomin and camphorin from the seeds of Cinnamomum camphora has been developed. Cinnamomin is a type II ribosome-inactivating protein (RIP), while camphorin is a type I RIP. Cinnamomin was purified by a single step of acid-treated Sepharose 4B affinity chromatography instead of gel filtration. Camphorin was purified by anion-exchange chromatography and gel filtration successively from the eluant not retained by the affinity column. Using this improved method, 620 mg of cinnamomin and 14.2 mg of camphorin were obtained from 500 g of wet seed, while only 10.6 mg of cinnamomin and 4.7 mg of camphorin were obtained by the previous method. Cinnamomin and camphorin purified by this method were homogeneous in SDS-denatured polyacrylamide gel electrophoresis. These two RIPs are multifunctional proteins. The assay of the enzymatic activities of cinnamomin and camphorin showed that both of them exhibit RNA N-glycosidase and supercoil-dependent endonuclease activities, while camphorin also exhibits superoxide dismutase activity.
Li
X D
XD
Shanghai Institute of Biochemistry, Academia Sinica, China.
Chen
W F
WF
Liu
W Y
WY
Wang
G H
GH
D020418:Algal Proteins; D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000187:drug effects; D009699:N-Glycosyl Hydrolases; D010940:Plant Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D010945:Plants, Edible / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D054790:Ribosome Inactivating Proteins, Type 2; D012270:Ribosomes / Q000187:drug effects*; D012639:Seeds / Q000737:chemistry
D020418:Algal Proteins; D010940:Plant Proteins; D011506:Proteins; D054790:Ribosome Inactivating Proteins, Type 2; C104406:camphorin protein, Cinnamomum camphora; C061930:cinnamomin, Phytophthora cinnamomi; D009699:N-Glycosyl Hydrolases