PDBsum entry 4fkd

Transferase

PDB id: 4fkd

Contents

Protein chain

65 a.a.

Metals

_ZN ×2

Waters ×130

PDB id:

4fkd

Name:

Transferase

Title:

Identification of the activator binding residues in the second cysteine-rich regulatory domain of protein kinasE C theta

Structure:

Protein kinasE C theta type. Chain: a. Synonym: npkc-theta. Engineered: yes

Source:

Mus musculus. Mouse. Organism_taxid: 10090. Gene: prkcq, pkcq. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.63Å R-factor: 0.173 R-free: 0.204

Authors:

G.M.Rahman,S.Shanker,N.E.Lewin,B.V.V.Prasad,P.M.Blumberg,J.Das

Key ref:

G.M.Rahman et al. (2013). Identification of the activator-binding residues in the second cysteine-rich regulatory domain of protein kinase Cθ (PKCθ). Biochem J, 451, 33-44. PubMed id: 23289588 DOI: 10.1042/BJ20121307

Date:

13-Jun-12 Release date: 23-Jan-13

Protein chain

Q02111  (KPCT_MOUSE) -  Protein kinase C theta type from Mus musculus

Seq: 707 a.a.

Struc: 65 a.a.*

* PDB and UniProt seqs differ at 14 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.7.11.13 - protein kinase C.
• Reaction:
  1. L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] + ADP + H⁺
  2. L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] + ADP + H⁺

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

reference

DOI no: 10.1042/BJ20121307

Biochem J 451:33-44 (2013)

PubMed id: 23289588

Identification of the activator-binding residues in the second cysteine-rich regulatory domain of protein kinase Cθ (PKCθ).

G.M.Rahman, S.Shanker, N.E.Lewin, N.Kedei, C.S.Hill, B.V.Prasad, P.M.Blumberg, J.Das.

ABSTRACT

PKC (protein kinase C) θ is predominantly expressed in T-cells and is critically involved in immunity. Design of PKCθ-selective molecules to manage autoimmune disorders by targeting its activator-binding C1 domain requires the knowledge of its structure and the activator-binding residues. The C1 domain consists of twin C1 domains, C1A and C1B, of which C1B plays a critical role in the membrane translocation and activation of PKCθ. In the present study we determined the crystal structure of PKCθC1B to 1.63 Å (1 Å=0.1 nm) resolution, which showed that Trp253 at the rim of the activator-binding pocket was orientated towards the membrane, whereas in PKCδC1B the homologous tryptophan residue was orientated away from the membrane. This particular orientation of Trp253 affects the size of the activator-binding pocket and the membrane affinity. To further probe the structural constraints on activator-binding, five residues lining the activator-binding site were mutated (Y239A, T243A, W253G, L255G and Q258G) and the binding affinities of the PKCθC1B mutants were measured. These mutants showed reduced binding affinities for phorbol ester [PDBu (phorbol 12,13-dibutyrate)] and diacylglycerol [DOG (sn-1,2-dioctanoylglycerol), SAG (sn-1-stearoyl 2-arachidonyl glycerol)]. All five full-length PKCθ mutants exhibited reduced phorbol-ester-induced membrane translocation compared with the wild-type. These results provide insights into the PKCθ activator-binding domain, which will aid in future design of PKCθ-selective molecules.