PDBsum entry: 1xjd

Transferase

PDB id: 1xjd

Contents

Protein chain

282 a.a. *

Ligands

STU

Waters ×115

* Residue conservation analysis

PDB id:

1xjd

Name:

Transferase

Title:

Crystal structure of pkc-theta complexed with staurosporine at 2a resolution

Structure:

Protein kinasE C, theta type. Chain: a. Fragment: residues 362-706: includes protein kinase domain (residues 380-634). Synonym: pkc-theta, npkc-theta. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: pkc. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Other_details: the sequence of this peptide can be found naturally in homo sapiens (human).

Resolution:

2.00Å R-factor: 0.201 R-free: 0.216

Authors:

Z.B.Xu

Key ref:

Z.B.Xu et al. (2004). Catalytic domain crystal structure of protein kinase C-theta (PKCtheta). J Biol Chem, 279, 50401-50409. PubMed id: 15364937 DOI: 10.1074/jbc.M409216200

Date:

23-Sep-04 Release date: 19-Oct-04

Protein chain

Q04759  (KPCT_HUMAN) -  Protein kinase C theta type

Seq: 706 a.a.

Struc: 282 a.a.*

* PDB and UniProt seqs differ at 9 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.7.11.13 - Protein kinase C.
• Reaction: ATP + a protein = ADP + a phosphoprotein

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

 Gene Ontology (GO) functional annotation 

• Biological process: protein amino acid phosphorylation (1 term)
• Biochemical function: protein kinase activity (3 terms)

reference

DOI no: 10.1074/jbc.M409216200

J Biol Chem 279:50401-50409 (2004)

PubMed id: 15364937

Catalytic domain crystal structure of protein kinase C-theta (PKCtheta).

Z.B.Xu, D.Chaudhary, S.Olland, S.Wolfrom, R.Czerwinski, K.Malakian, L.Lin, M.L.Stahl, D.Joseph-McCarthy, C.Benander, L.Fitz, R.Greco, W.S.Somers, L.Mosyak.

ABSTRACT

A member of the novel protein kinase C (PKC) subfamily, PKC, is an essential component of the T cell synapse and is required for optimal T cell activation and interleukin-2 production. Selective involvement of PKC in TCR signaling makes this enzyme an attractive therapeutic target in T cell-mediated disease processes. In this report we describe the crystal structure of the catalytic domain of PKC at 2.0-A resolution. Human recombinant PKC kinase domain was expressed in bacteria as catalytically active phosphorylated enzyme and co-crystallized with its subnanomolar, ATP site inhibitor staurosporine. The structure follows the classic bilobal kinase fold and shows the enzyme in its active conformation and phosphorylated state. Inhibitory interactions between conserved features of staurosporine and the ATP-binding cleft are accompanied by closing of the glycine-rich loop, which also maintains an inhibitory arrangement by blocking the phosphate recognition subsite. The two major phosphorylation sites, Thr-538 in the activation loop and Ser-695 in the hydrophobic motif, are both occupied in the structure, playing key roles in stabilizing active conformation of the enzyme and indicative of PKC autocatalytic phosphorylation and activation during bacterial expression. The PKC-staurosporine complex represents the first kinase domain crystal structure of any PKC isotypes to be determined and as such should provide valuable insight into PKC specificity and into rational drug design strategies for PKC selective leads.

Selected figure(s)

Figure 2.
FIG. 2. Overall structure of the PKC -staurosporine complex and comparison with the structure of the PKA-staurosporine-PKI complex. A, ribbon representation of the PKC kinase domain structure. The N-lobe is cyan and the C-lobe is blue. The glycine-rich loop, activation loop, and HM segment are highlighted in red. Staurosporine and phosphorylated residues are shown in stick representation. B, superposition of PKC -staurosporine (color coding is the same as described for A) and PKA-staurosporine-PKI structures (yellow ribbons and sticks; PDB code: 1STC [PDB] (20)). Phosphorylated sites in PKA are also shown. Structures were aligned using the central helices from the C-lobe. Both staurosporine-bound kinases display intermediate lobe structures with conformational differences in the glycine-rich loop.

Figure 4.
FIG. 4. The activation loop and helix C. The ribbon drawing of PKC (N-lobe in cyan, C-lobe in blue) highlights key structural motifs at the catalytic cleft. Selected residues are in stick representation. Dashed red lines depict specific hydrogen-bonding interactions of the essential Thr-538(P) phosphate with cationic pocket (Arg-503, Lys-527) and of the C helix with catalytic Lys-409 and activation loop (Glu-528).

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2004, 279, 50401-50409) copyright 2004.

Figures were selected by the author.