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嗜酸丛毛单胞菌天然及化学修饰的阴离子选择性孔蛋白的结晶及初步X射线晶体学研究

Crystallization and preliminary X-ray crystallographic studies of the native and chemically modified anion-selective porin from Comamonas acidovorans.

作者信息

Zeth K, Schnaible V, Przybylski M, Welte W, Diederichs K, Engelhardt H

机构信息

Max-Planck-Institut für Biochemie, Am Klopferspitz 18a, D-82152 Martinsried, Germany.

出版信息

Acta Crystallogr D Biol Crystallogr. 1998 Jul 1;54(Pt 4):650-3. doi: 10.1107/s0907444997018854.

Abstract

Omp32, the strongly anion-selective porin from Comamonas acidovorans, has been crystallized. Two crystal forms were observed, both of which belong to space group R3, but exhibit different cell dimensions a = b = 106.7, c = 140.6 A (crystal form I) and a = b = 87.1, c = 135.3 A (crystal form II) with one trimer per asymmetric unit. The crystals diffract to 2.2 and 2.3 A resolution, respectively. Omp32 was chemically modified by introducing negative charges through succinylation. The number and positions of the individual modifications were determined using mass spectrometry and X-ray crystallography. Chemically modified porins yielded crystals of a third form, also of space group R3 but with cell constants of a = b = 109.3 and c = 263.2 A (crystal form III), showing a virtually doubled c axis. Crystals of form III diffract to 3.5 A resolution.

摘要

嗜酸丛毛单胞菌中具有强阴离子选择性的孔蛋白Omp32已被结晶。观察到两种晶体形式,它们都属于R3空间群,但具有不同的晶胞尺寸,a = b = 106.7,c = 140.6 Å(晶体形式I)和a = b = 87.1,c = 135.3 Å(晶体形式II),每个不对称单元中有一个三聚体。这些晶体的衍射分辨率分别为2.2 Å和2.3 Å。通过琥珀酰化引入负电荷对Omp32进行化学修饰。使用质谱和X射线晶体学确定各个修饰的数量和位置。化学修饰的孔蛋白产生了第三种晶体形式,同样属于R3空间群,但晶胞常数为a = b = 109.3和c = 263.2 Å(晶体形式III),显示c轴几乎加倍。晶体形式III的晶体衍射分辨率为3.5 Å。

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