Purification, crystallization and preliminary X-ray analysis of two crystal forms of ribonuclease Sa3
| Autor: | Lubica Urbanikova, Vladena Hlinková, Jozef Sevcik, Daniela Krajcikova |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
biology
Strain (chemistry) Protein Conformation RNase P Chemistry Streptomyces aureofaciens General Medicine Crystallography X-Ray biology.organism_classification medicine.disease_cause law.invention Isoenzymes Crystallography Tetragonal crystal system Ribonucleases Protein structure Structural Biology law biology.protein medicine Ribonuclease Crystallization Escherichia coli |
| Zdroj: | Acta Crystallographica Section D Biological Crystallography. 57:737-739 |
| ISSN: | 0907-4449 |
| DOI: | 10.1107/s0907444901003456 |
| Popis: | RNase Sa3 produced by Streptomyces aureofaciens strain CCM 3239 belongs to the T1 family of microbial ribonucleases. It is closely related both to RNase Sa, studied in detail earlier, and to RNase Sa2 produced by the same microorganism. The most important property of RNase Sa3 is the relatively high cytotoxic activity, which was not observed for RNase Sa and Sa2. Recombinant RNase Sa3 was overexpressed in Escherichia coli and purified to high homogeneity. The hanging-drop vapour-diffusion method was used for crystallization. The two crystal forms are trigonal P3(1)21 and tetragonal P4(1)2(1)2, with unit-cell parameters a = b = 64.7, c = 69.6 A, gamma = 120 degrees and a = b = 34.0, c = 147.2 A, respectively. They diffract to 2.0 and to 1.7 A resolution, respectively, using synchrotron radiation. The asymmetric units of crystal forms I and II contain one molecule of the enzyme, which corresponds to V(M) = 3.8 A(3) Da(-1) with a solvent content of 68% and V(M) = 1.9 A(3) Da(-1) with a solvent content of 37%, respectively. |
| Databáze: | OpenAIRE |
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