PDBsum entry 3geq

Transferase

PDB id: 3geq

Contents

Protein chains

272 a.a. *

Ligands

PP2 ×2

Waters ×221

* Residue conservation analysis

PDB id:

3geq

Name:

Transferase

Title:

Structural basis for the chemical rescue of src kinase activity

Structure:

Proto-oncogene tyrosine-protein kinase src. Chain: a, b. Fragment: kinase domain (unp residues 251-533). Synonym: pp60c-src, p60-src, c-src. Engineered: yes. Mutation: yes

Source:

Gallus gallus. Bantam,chickens. Organism_taxid: 9031. Gene: src. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.20Å R-factor: 0.226 R-free: 0.273

Authors:

K.E.Muratore,M.A.Seeliger,Z.Wang,D.Fomina,J.Neiswinger,J.J.Havranek, D.Baker,J.Kuriyan,P.A.Cole

Key ref:

K.E.Muratore et al. (2009). Comparative analysis of mutant tyrosine kinase chemical rescue. Biochemistry, 48, 3378-3386. PubMed id: 19260709

Date:

25-Feb-09 Release date: 28-Apr-09

Protein chains

P00523  (SRC_CHICK) -  Proto-oncogene tyrosine-protein kinase Src from Gallus gallus

Seq: 533 a.a.

Struc: 272 a.a.*

* PDB and UniProt seqs differ at 4 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.7.10.2 - non-specific protein-tyrosine kinase.
• Reaction: L-tyrosyl-[protein] + ATP = O-phospho-L-tyrosyl-[protein] + ADP + H⁺

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

Biochemistry 48:3378-3386 (2009)

PubMed id: 19260709

Comparative analysis of mutant tyrosine kinase chemical rescue.

K.E.Muratore, M.A.Seeliger, Z.Wang, D.Fomina, J.Neiswinger, J.J.Havranek, D.Baker, J.Kuriyan, P.A.Cole.

ABSTRACT

Protein tyrosine kinases are critical cell signaling enzymes. These enzymes have a highly conserved Arg residue in their catalytic loop which is present two residues or four residues downstream from an absolutely conserved Asp catalytic base. Prior studies on protein tyrosine kinases Csk and Src revealed the potential for chemical rescue of catalytically deficient mutant kinases (Arg to Ala mutations) by small diamino compounds, particularly imidazole; however, the potency and efficiency of rescue was greater for Src. This current study further examines the structural and kinetic basis of rescue for mutant Src as compared to mutant Abl tyrosine kinase. An X-ray crystal structure of R388A Src revealed the surprising finding that a histidine residue of the N-terminus of a symmetry-related kinase inserts into the active site of the adjacent Src and mimics the hydrogen-bonding pattern seen in wild-type protein tyrosine kinases. Abl R367A shows potent and efficient rescue more comparable to Src, even though its catalytic loop is more like that of Csk. Various enzyme redesigns of the active sites indicate that the degree and specificity of rescue are somewhat flexible, but the overall properties of the enzymes and rescue agents play an overarching role. The newly discovered rescue agent 2-aminoimidazole is about as efficient as imidazole in rescuing R/A Src and Abl. Rate vs pH studies with these imidazole analogues suggest that the protonated imidazolium is the preferred form for chemical rescue, consistent with structural models. The efficient rescue seen with mutant Abl points to the potential of this approach to be used effectively to analyze Abl phosphorylation pathways in cells.