Expression, purification, crystallization and preliminary X-ray crystallographic analysis of tomato β-galactosidase 4
| Autor: | Toshiji Tada, Masahiro Eda, Megumi Ishimaru |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Molecular Sequence Data
Biophysics Crystallography X-Ray Biochemistry Pichia pastoris law.invention Research Communications chemistry.chemical_compound Solanum lycopersicum Structural Biology law PEG ratio Genetics Amino Acid Sequence Crystallization biology Condensed Matter Physics biology.organism_classification beta-Galactosidase Yeast Solvent Crystallography Monomer chemistry Recombinant DNA Orthorhombic crystal system Electrophoresis Polyacrylamide Gel |
| Popis: | Plant β-galactosidases play important roles in carbohydrate-reserve mobilization, cell-wall expansion and degradation, and turnover of signalling molecules during ripening. Tomato β-galactosidase 4 (TBG4) not only has β-galactosidase activity but also has exo-β-(1,4)-galactanase activity, and prefers β-(1,4)-galactans longer than pentamers as its substrates; most other β-galactosidases only have the former activity. Recombinant TBG4 protein expressed in the yeastPichia pastoriswas crystallized by the sitting-drop vapour-diffusion method using PEG 10 000 as a precipitant. The crystals belonged to the orthorhombic space groupP212121, with unit-parametersa= 92.82,b= 96.30,c= 159.26 Å, and diffracted to 1.65 Å resolution. Calculation of the Matthews coefficient suggested the presence of two monomers per asymmetric unit (VM= 2.2 Å3 Da−1), with a solvent content of 45%. |
| Databáze: | OpenAIRE |
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