Identification, cDNA cloning, expression, crystallization and preliminary X-ray analysis of an exceptionally halotolerant carbonic anhydrase from Dunaliella salina.

Premkumar L; Greenblatt HM; Bageshwar UKSavchenko TGokhman IZamir ASussman JL

首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >Identification, cDNA cloning, expression, crystallization and preliminary X-ray analysis of an exceptionally halotolerant carbonic anhydrase from Dunaliella salina.

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Identification, cDNA cloning, expression, crystallization and preliminary X-ray analysis of an exceptionally halotolerant carbonic anhydrase from Dunaliella salina.

机译：识别、cDNA克隆,表达,结晶和初步的x射线分析异常halotolerant碳酸酐酶杜氏。

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An extracellular alpha-type carbonic anhydrase (dCAII) from the salt-tolerant alga Dunaliella salina differs from its mesophilic counterparts in remaining active from zero to multimolar salt concentrations. To gain insight into the outstanding salt tolerance of dCAII, the enzyme was functionally overexpressed in Escherichia coli, purified by affinity chromatography and crystallized by the hanging-drop method. The crystals belonged to space group P2(1), with unit-cell parameters a = 47.0, b = 119.9, c = 58.5 A, beta = 94.2 degrees. Data from a single crystal were collected to 2.4 A resolution under cryogenic conditions (120 K) using an R-AXIS IV(++) detector mounted on a Rigaku RU-H3R rotating-anode generator. The asymmetric unit contains two molecules of the protein, which corresponds to V(M) = 2.65 A(3) Da(-1) and a solvent content of 52.7%.

机译：一个细胞外alpha-type碳酸酐酶从耐盐藻杜氏盐藻(dCAII)赛利娜不同于嗜中温同行活跃在从零到multimolar盐浓度。优秀盐dCAII公差,酶功能在大肠吗杆菌、亲和色谱法和净化悬滴结晶的方法。水晶属于空间群P2 (1)晶胞参数= 47.0,b = 119.9, c =58.5,β= 94.2度。收集晶体分辨率在2.4低温条件下(120 K)使用一个轴IV(+ +)探测器安装在Rigaku RU-H3R旋转阳极生成器。包含两个分子的蛋白质,对应于V (M) = 2.65 (3) Da(1)和一个溶剂含量的52.7%。

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《Acta crystallographica.Section D. Biological crystallography》 |2003年第6期|1084-1086|共3页
作者
Premkumar L; Greenblatt HM; Bageshwar UKSavchenko TGokhman IZamir ASussman JL;
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作者单位

Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.;

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正文语种英语
中图分类化学;
关键词
Carbonic Anhydrases; Affinity Chromatography; X-ray analysisSaltsDunaliella salinaX-raysCrystallization;

机译：碳酸脱水酶;亲和色谱法;x射线salinaX-raysCrystallization;

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Identification, cDNA cloning, expression, crystallization and preliminary X-ray analysis of an exceptionally halotolerant carbonic anhydrase from Dunaliella salina.

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