Second-generation octarellins: two new de novo (β/α)8 polypeptides designed for investigating the influence of β-residue packing on the α/β-barrel structure stability
| Autor: | Jean Marie Ruysschaert, Alain Poncin, Alain Lamproye, Gisèle Préaux, Erik Goormaghtigh, Ruben Abagyan, Benoît Moreau, Joseph Martial, Annick Houbrechts, Véronique Mainfroid, Karine Goraj |
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| Rok vydání: | 1995 |
| Předmět: |
Protein Denaturation
Circular dichroism Dimer Molecular Sequence Data Protozoan Proteins Bioengineering Protein Engineering Biochemistry Protein Structure Secondary Fungal Proteins chemistry.chemical_compound Protein structure Bacterial Proteins Spectroscopy Fourier Transform Infrared Escherichia coli Genes Synthetic Urea Amino Acid Sequence Beta (finance) Molecular Biology Protein secondary structure Peptide sequence Gel electrophoresis Base Sequence Circular Dichroism Recombinant Proteins Crystallography Beta barrel Solubility chemistry Chromatography Gel Electrophoresis Polyacrylamide Gel Peptides Ultracentrifugation Biotechnology |
| Zdroj: | "Protein Engineering, Design and Selection". 8:249-259 |
| ISSN: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/8.3.249 |
| Popis: | The sequence of octarellin I, the first de novo (beta/alpha)8 polypeptide, was revised according to several criteria, among others the symmetry of the sequence, beta-residue volume and hydrophobicity, and charge distribution. These considerations and the overall conclusions drawn from the first design led to two new sequences, corresponding to octarellins II and III. Octarellin II retains perfect 8-fold symmetry. Octarellin III has the same sequence as octarellin II, except for the beta-strands which exhibit a 4-fold symmetry. The two proteins were produced in Escherichia coli. Infrared and CD spectral analyses of octarellins II and III reveal a high secondary structure content. Non-denaturing gel electrophoresis, molecular sieve chromatography and analytical ultracentrifugation suggest that both of these second-generation artificial polypeptides exist as a mixture of a monomer and a dimer form. Octarellins II and III are at least 10 times more soluble than octarellin I. Urea-induced unfolding followed by fluorescence emission suggests that the tryptophan residues, designed to be buried in the (beta/alpha)8, are indeed packed in the hydrophobic core of both proteins. However, octarellin III displays a higher stability towards urea denaturation, indicating that introducing 4-fold symmetry into the beta-barrel might be important for stability of the overall folding. |
| Databáze: | OpenAIRE |
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