PDBsum entry 1r3c

Transferase

PDB id: 1r3c

Contents

Protein chain

349 a.a. *

Metals

_MG ×2

Waters ×268

* Residue conservation analysis

PDB id:

1r3c

Name:

Transferase

Title:

The structure of p38alpha c162s mutant

Structure:

Mitogen-activated protein kinase 14. Chain: a. Synonym: mitogen-activated protein kinase p38alpha, map kinase p38alpha, cytokine suppressive anti-inflammatory drug binding protein, csaid binding protein, csbp, max-interacting protein 2, map kinase mxi2, sapk2a. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: mapk14. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.

Resolution:

2.00Å R-factor: 0.200 R-free: 0.224

Authors:

S.B.Patel,P.M.Cameron,B.Frantz-Wattley,E.O'Neill,J.W.Becker,G.Scapin

Key ref:

S.B.Patel et al. (2004). Lattice stabilization and enhanced diffraction in human p38 alpha crystals by protein engineering. Biochim Biophys Acta, 1696, 67-73. PubMed id: 14726206

Date:

01-Oct-03 Release date: 20-Jan-04

Protein chain

Q16539  (MK14_HUMAN) -  Mitogen-activated protein kinase 14 from Homo sapiens

Seq: 360 a.a.

Struc: 349 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.2.7.11.24 - mitogen-activated protein kinase.
• 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

Biochim Biophys Acta 1696:67-73 (2004)

PubMed id: 14726206

Lattice stabilization and enhanced diffraction in human p38 alpha crystals by protein engineering.

S.B.Patel, P.M.Cameron, B.Frantz-Wattley, E.O'Neill, J.W.Becker, G.Scapin.

ABSTRACT

Mitogen-activated protein (MAP) kinase p38 alpha is activated in response to environmental stress and cytokines, and plays a significant role in inflammatory responses. For these reasons, it is an important target for the treatment of a wide range of inflammatory and autoimmune diseases. The crystals of p38 alpha that we obtained by published procedures were usually small, quite mosaic, and difficult to reproduce and thus posed a difficulty for the intensive high-resolution studies required for a structure-guided drug discovery approach. Based on crystallographic and biochemical evidences, we prepared a single point mutation of a surface cysteine (C162S) and found that it prevents aggregation and improves the homogeneity and stability of the enzyme. This mutation also facilitates the crystallization process and increases the diffracting power of p38 alpha crystals. Surprisingly, we found that the mutation induces a change in the conformation of a nearby surface loop resulting in stronger lattice interactions, consistent with the improved crystal quality. The mutant protein, because of its improved stability and strengthened lattice interactions, thus provides a significantly improved reagent for use in structure-based drug design for this important disease target.