1KSW

Structure of Human c-Src Tyrosine Kinase (Thr338Gly Mutant) in Complex with N6-benzyl ADP

Summary for 1KSW

• Entry DOI: 10.2210/pdb1ksw/pdb
• Related: 1FMK 2SRC
• Descriptor: PROTO-ONCOGENE TYROSINE-PROTEIN KINASE SRC, N6-BENZYL ADENOSINE-5'-DIPHOSPHATE (3 entities in total)
• Functional Keywords: sh3, sh2, kinase, bump hole, bump-hole, chemical genetics, orthogonal substrate, atp, transferase
• Biological source: Homo sapiens (human)
• Cellular location: Cell membrane: P12931
• Total number of polymer chains: 1
• Total formula weight: 52182.84

Authors

Witucki, L.A.,Huang, X.,Shah, K.,Liu, Y.,Kyin, S.,Eck, M.J.,Shokat, K.M. (deposition date: 2002-01-14, release date: 2002-02-27, Last modification date: 2023-11-15)

Primary citation

Witucki, L.A.,Huang, X.,Shah, K.,Liu, Y.,Kyin, S.,Eck, M.J.,Shokat, K.M.
Mutant tyrosine kinases with unnatural nucleotide specificity retain the structure and phospho-acceptor specificity of the wild-type enzyme.
Chem.Biol., 9:25-33, 2002

PubMed Abstract: The direct substrates of one protein kinase in a cell can be identified by mutation of the ATP binding pocket to allow an unnatural ATP analog to be accepted exclusively by the engineered kinase. Here, we present structural and functional assessment of peptide specificity of mutant protein kinases with unnatural ATP analogs. The crystal structure (2.8 A resolution) of c-Src (T338G) with N(6)-(benzyl) ADP bound shows that the creation of a unique nucleotide binding pocket does not alter the phospho-acceptor binding site of the kinase. A panel of optimal peptide substrates of defined sequence, as well as a degenerate peptide library, was utilized to assess the phospho-acceptor specificity of the engineered "traceable" kinases. The specificity profiles for the mutant kinases were found to be identical to those of their wild-type counterparts.

PubMed: 11841936
DOI: 10.1016/S1074-5521(02)00091-1

Experimental method

X-RAY DIFFRACTION (2.8 Å)