Expression, purification, crystallization and preliminary crystallographic analysis of human Pim-1 kinase
- Department of Immunology and Inflammation, Boehringer Ingelheim Pharmaceuticals Inc., Research and Development, 175 Briar Ridge Road, Ridgefield, CT 06877 (United States)
- Department of Medicinal Chemistry, Boehringer Ingelheim Pharmaceuticals Inc., Research and Development, 175 Briar Ridge Road, Ridgefield, CT 06877 (United States)
Pim kinases, belong to a distinctive serine/threonine protein-kinase family and are involved in cytokine-induced signal transduction and the development of lymphoid malignancies. Human Pim-1 kinase has been cloned, expressed and crystallized 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 Å resolution and belong to space group P6{sub 5}, with unit-cell parameters a = b = 95.9, c = 80.0 Å, β = 120° and one molecule per asymmetric unit.
- OSTI ID:
- 22356084
- Journal Information:
- Acta Crystallographica. Section F, Vol. 61, Issue Pt 1; Other Information: PMCID: PMC1952393; PMID: 16508102; PUBLISHER-ID: en5073; OAI: oai:pubmedcentral.nih.gov:1952393; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
Similar Records
Purification, crystallization and preliminary X-ray diffraction analysis of the kinase domain of human tousled-like kinase 2
Biochemical and molecular analysis of a transmembrane protein kinase from Arabidopsis thaliana