Qian Kevin C, Studts Joey, Wang Lian, Barringer Kevin, Kronkaitis Anthony, Peng Charline, Baptiste Alistair, LaFrance Roger, Mische Sheenah, Farmer Bennett
Department of Medicinal Chemistry, Boehringer Ingelheim Pharmaceuticals Inc., Research and Development, Ridgefield, CT 06877, USA.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Jan 1;61(Pt 1):96-9. doi: 10.1107/S1744309104029963. Epub 2004 Dec 2.
Pim kinases, including Pim-1, Pim-2 and Pim-3, belong to a distinctive serine/threonine protein-kinase family. They are involved in cytokine-induced signal transduction and the development of lymphoid malignancies. Their kinase domains are highly homologous to one another, but share low sequence identity to other kinases. Specifically, there are two proline residues in the conserved hinge-region sequence ERPXPX separated by a residue that is non-conserved among Pim kinases. Full-length human Pim-1 kinase (1-313) was cloned and expressed in Escherichia coli as a GST-fusion protein and truncated to Pim-1 (14-313) by thrombin digestion during purification. The Pim-1 (14-313) protein was purified to high homogeneity and monodispersity. This protein preparation yielded small crystals in the initial screening and large crystals after optimization. The large crystals of apo Pim-1 enzyme diffracted to 2.1 A resolution and belong to space group P6(5), with unit-cell parameters a = b = 95.9, c = 80.0 A, beta = 120 degrees and one molecule per asymmetric unit.
Pim激酶，包括Pim-1、Pim-2和Pim-3，属于一个独特的丝氨酸/苏氨酸蛋白激酶家族。它们参与细胞因子诱导的信号转导以及淋巴恶性肿瘤的发展。它们的激酶结构域彼此高度同源，但与其他激酶的序列同一性较低。具体而言，在保守的铰链区序列ERPXPX中有两个脯氨酸残基，中间隔着一个在Pim激酶中不保守的残基。全长人Pim-1激酶（1-313）被克隆并在大肠杆菌中作为GST融合蛋白表达，并在纯化过程中通过凝血酶消化截短为Pim-1（14-313）。Pim-1（14-313）蛋白被纯化至高纯度和单分散性。该蛋白制剂在初始筛选中产生了小晶体，优化后得到了大晶体。无配体Pim-1酶的大晶体衍射分辨率达到2.1 Å，属于空间群P6(5)，晶胞参数a = b = 95.9，c = 80.0 Å，β = 120°，每个不对称单元中有一个分子。
