Purification and characterization of novel ribosome inactivating proteins, alpha- and beta-pisavins, from seeds of the garden pea Pisum sativum

被引:184
作者
Lam, SSL [1 ]
Wang, HX
Ng, TB
机构
[1] Chinese Univ Hong Kong, Fac Med, Dept Biochem, Shatin, Peoples R China
[2] China Agr Univ, Dept Microbiol, Beijing, Peoples R China
关键词
ribosome inactivating proteins; garden pea; pisavin;
D O I
10.1006/bbrc.1998.9764
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Two ribosome inactivating proteins designated alpha-and beta-pisavins were isolated from seeds of the garden pea Pisum sativum var. arvense Poir with a procedure involving affinity chromatography on Affi-gel Blue gel, immobilized metal ion affinity chromatography on Iminodiacetic acid-agarose, cation exchange chromatography on Resource-S, and gel filtration on Superose 12. alpha- and beta-pisavins are nonglycoproteins with a molecular weight of 20.5 kDa and 18.7 kDa respectively. The sequences of the fist sixty N-terminal amino acids of alpha- and beta-pisavins were identical. In isoelectric focusing these two proteins merged into one band with a pi greater than 9.3. Inhibition of protein synthesis by a rabbit reticulocyte lysate system was achieved at an IC50 of approximately 0.5 nM. Activity of the proteins toward tRNA was observed. The proteins acted on ribosomal RNA through its RNA N-glycosidase activity to release an Endo's fragment, and converted the conformation of DNA from supercoiled and circular forms into a linear form. (C) 1998 Academic Press.
引用
收藏
页码:135 / 142
页数:8
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