title: Cloning and expression of cDNA coding for bouganin.
abstract: Bouganin is a ribosome-inactivating protein that recently was isolated from Bougainvillea spectabilis Willd. In this work, the cloning and expression of the cDNA encoding for bouganin is described. From the cDNA, the amino-acid sequence was deduced, which correlated with the primary sequence data obtained by amino-acid sequencing on the native protein. Bouganin is synthesized as a pro-peptide consisting of 305 amino acids, the first 26 of which act as a leader signal while the 29 C-terminal amino acids are cleaved during processing of the molecule. The mature protein consists of 250 amino acids. Using the cDNA sequence encoding the mature protein of 250 amino acids, a recombinant protein was expressed, purified and characterized. The recombinant molecule had similar activity in a cell-free protein synthesis assay and had comparable toxicity on living cells as compared to the isolated native bouganin.
authors.affiliation: Tanox Pharma B.V., Amsterdam, the Netherlands; Dipartimento di Patologia Sperimentale, Università di Bologna, Bologna, Italy. mtdenhartog@hotmail.com
authors.forename: Marcel T
authors.initials: MT
authors.lastname: den Hartog
authors.forename: Chiara
authors.initials: C
authors.lastname: Lubelli
authors.forename: Louis
authors.initials: L
authors.lastname: Boon
authors.forename: Sijmie
authors.initials: S
authors.lastname: Heerkens
authors.forename: Antonio P
authors.initials: AP
authors.lastname: Ortiz Buijsse
authors.forename: Mark
authors.initials: M
authors.lastname: de Boer
authors.forename: Fiorenzo
authors.initials: F
authors.lastname: Stirpe
mesh_terms: D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D019684:Magnoliopsida / Q000737:chemistry*; D008969:Molecular Sequence Data; D009699:N-Glycosyl Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D054788:Ribosome Inactivating Proteins; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D009699:N-Glycosyl Hydrolases; D054788:Ribosome Inactivating Proteins