Purification, crystallization and preliminary X-ray data collection of the N-terminal domain of the 26S proteasome regulatory subunit p27 and its complex with the ATPase domain of Rpt5 from Mus musculus
| Autor: | Xue Yang, Wentao Diao, Hao Zhou |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Proteasome Endopeptidase Complex
Protein subunit ATPase Biophysics Crystallography X-Ray Biochemistry law.invention Mice Structural Biology law Genetics Molecule Animals Crystallization Adenosine Triphosphatases biology Resolution (electron density) Condensed Matter Physics Protein Structure Tertiary Solvent Crystallography Protein Subunits Proteasome Crystallization Communications Domain (ring theory) biology.protein |
| Zdroj: | Acta crystallographica. Section F, Structural biology communications. 70(Pt 5) |
| ISSN: | 2053-230X |
| Popis: | The protein 26S proteasome regulatory subunit p27 is one of the four chaperones that help in the assembly of the 19S regulatory particle (RP) of the 26S proteasome. In the present work, the N-terminus of p27 (residues 1–128) fromMus musculuswas cloned, expressed, purified and crystallized alone and in complex with the C-terminal ATPase domain of Rpt5 (residues 173–442). The crystals of p27(1–128)diffracted to 1.7 Å resolution and belonged to space groupP212121, with unit-cell parametersa= 26.79,b= 30.39,c= 145.06 Å. Resolution-dependent Matthews coefficient probability analysis suggested the presence of only one molecule per asymmetric unit, with 40.5% solvent content and aVMvalue of 2.02 Å3 Da−1. The crystal of the p27(1–128)–Rpt5(173–442)complex diffracted to 4 Å resolution and belonged to space groupP222, with unit-cell parametersa= 75.93,b= 76.08,c= 336.85 Å. The presence of four heterodimers in the asymmetric unit with 53.2% solvent content and aVMvalue of 2.63 Å3 Da−1or five heterodimers in the asymmetric unit with 41.5% solvent content and aVMvalue of 2.10 Å3 Da−1is assumed. |
| Databáze: | OpenAIRE |
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