Expression, purification, crystallization and preliminary crystallographic analysis of human Pim-1 kinase

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Kevin QianBennett T Farmer

Abstract

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.

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Citations

Mar 10, 2015·Pharmacology & Therapeutics·Noel A Warfel, Andrew S Kraft
Oct 9, 2012·Biochemical Pharmacology·Carmen Blanco-Aparicio, Amancio Carnero
Apr 12, 2013·Medicinal Research Reviews·Maja Narlik-GrassowAmancio Carnero
Oct 18, 2013·Expert Opinion on Therapeutic Patents·Gubbi M AruneshVellarkad N Viswanadhan
Jan 27, 2010·Expert Opinion on Therapeutic Patents·Tina Morwick
May 25, 2012·Carcinogenesis·Maja Narlik-GrassowAmancio Carnero
Nov 5, 2004·The Journal of Biological Chemistry·Kevin C QianBennett Farmer
Dec 12, 2020·Molecular Cancer Therapeutics·Rachel K Toth, Noel A Warfel
Aug 29, 2012·Journal of Medicinal Chemistry·Denis DryginDavid M Ryckman

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