A RECOMBINANT RIBOSOME-INACTIVATING PROTEIN FROM THE PLANT PHYTOLACCA-DIOICA L. PRODUCED FROM A SYNTHETIC GENE

Citation
Fd. Blanco et al., A RECOMBINANT RIBOSOME-INACTIVATING PROTEIN FROM THE PLANT PHYTOLACCA-DIOICA L. PRODUCED FROM A SYNTHETIC GENE, FEBS letters, 437(3), 1998, pp. 241-245
Citations number
21
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,"Cell Biology",Biophysics
Journal title
FEBS letters → ACNP
ISSN journal
0014-5793
Volume
437
Issue
3
Year of publication
1998
Pages
241 - 245
Database
ISI
SICI code
0014-5793(1998)437:3<241:ARRPFT>2.0.ZU;2-6
Abstract
Phytolacca dioica L. leaves produce at least two type-I ribosome-inact ivating proteins. Each polypeptide chain is subjected to different pos t-translational modifications giving rise to PD-L1 and PD-L2, and PD-L 3 and PD-L4, each polypeptide pair having the same primary structure. With the aim of exploiting the cytotoxic properties of these proteins as potential biological phytodrugs, a gene encoding PD-L4 was designed based on criteria expected to maximize the translation efficiency in tomato. The gene was constructed from 18 oligonucleotides and prelimin arily expressed in Escherichia coli, using the T7 promoter system. The protein produced was insoluble and accumulated in inclusion bodies to about 300 mg/l of culture. Ribosome-inactivating activity was generat ed by controlled oxidation of the reduced and denatured protein. The r ecombinant protein was indistinguishable from natural PD-L4 as isolate d from leaves of Phytolacca dioica, in both catalytic activity and pri mary structure. (C) 1998 Federation of European Biochemical Societies.