|Reference : Synthesis, purification and initial structural characterization of octarellin, a de n...|
|Scientific journals : Article|
|Life sciences : Biochemistry, biophysics & molecular biology|
|Synthesis, purification and initial structural characterization of octarellin, a de novo polypeptide modelled on the alpha/beta-barrel proteins|
|Goraj, Karine [> > > >]|
|Renard, André [ > > ]|
|Martial, Joseph [Université de Liège - ULg > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire >]|
|Oxford University Press|
|Yes (verified by ORBi)|
|[en] Amino Acid Sequence ; Automatic Data Processing ; Base Sequence ; Circular Dichroism ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Molecular Sequence Data ; *Peptides/genetics ; Plasmids ; Protein Conformation ; *Protein Engineering|
|[en] We have attempted to construct an artificial polypeptide that folds like the eight-stranded parallel beta-barrel structures. Our approach consists of repeating eight times a unit peptide designed to adopt a 'beta-strand/alpha-helix' pattern. A first 'test' sequence for this structural unit was deduced from a series of parameters defined after an analysis of three natural alpha/beta-barrel proteins and including principally the lengths of the secondary structure elements, the alpha/beta packing and the fitting on average Garnier profiles. The gene encoding this structural unit was synthesized, cloned and expressed in Escherichia coli either as a monomer or as direct repeats of 2-12 units. Preliminary structural characterization of the 7-, 8- and 9-fold unit polypeptides by circular dichroism measurements indicates the presence of the predicted amount of alpha-helix in the three proteins. Further analysis by urea-gradient gel electrophoresis demonstrates that, in the conditions tested, only the 8-fold unit polypeptide forms a compact structure through a cooperative and rapid two-state folding transition involving long-range molecular interactions.|
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