3SLS

Crystal Structure of human MEK-1 kinase in complex with UCB1353770 and AMPPNP

Summary for 3SLS

• Entry DOI: 10.2210/pdb3sls/pdb
• Descriptor: Dual specificity mitogen-activated protein kinase kinase 1, PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER, MAGNESIUM ION, ... (5 entities in total)
• Functional Keywords: serine-threonine kinase, signalling, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• Biological source: Homo sapiens (human)
• Cellular location: Cytoplasm, cytoskeleton, centrosome: Q02750
• Total number of polymer chains: 2
• Total formula weight: 69960.33

Authors

Meier, C.,Ceska, T.A. (deposition date: 2011-06-25, release date: 2012-02-29, Last modification date: 2024-02-28)

Primary citation

Meier, C.,Brookings, D.C.,Ceska, T.A.,Doyle, C.,Gong, H.,McMillan, D.,Saville, G.P.,Mushtaq, A.,Knight, D.,Reich, S.,Pearl, L.H.,Powell, K.A.,Savva, R.,Allen, R.A.
Engineering human MEK-1 for structural studies: A case study of combinatorial domain hunting.
J.Struct.Biol., 177:329-334, 2012

PubMed Abstract: Structural biology studies typically require large quantities of pure, soluble protein. Currently the most widely-used method for obtaining such protein involves the use of bioinformatics and experimental methods to design constructs of the target, which are cloned and expressed. Recently an alternative approach has emerged, which involves random fragmentation of the gene of interest and screening for well-expressing fragments. Here we describe the application of one such fragmentation method, combinatorial domain hunting (CDH), to a target which historically was difficult to express, human MEK-1. We show how CDH was used to identify a fragment which covers the kinase domain of MEK-1 and which expresses and crystallizes significantly better than designed expression constructs, and we report the crystal structure of this fragment which explains some of its superior properties. Gene fragmentation methods, such as CDH, thus hold great promise for tackling difficult-to-express target proteins.

PubMed: 22245778
DOI: 10.1016/j.jsb.2012.01.002

Experimental method

X-RAY DIFFRACTION (2.3 Å)