PDBsum entry 3vug

Transferase/transferase inhibitor

PDB id: 3vug

Contents

Protein chain

356 a.a.

Ligands

PRO-LYS-ARG-PRO-THR-THR-LEU-ASN-LEU-PHE

SO4 ×4

Waters ×28

PDB id:

3vug

Name:

Transferase/transferase inhibitor

Title:

Crystal structure of a cysteine-deficient mutant m2 in map kinase jnk1

Structure:

Mitogen-activated protein kinase 8. Chain: a. Fragment: kinase domain, unp residues 1-364. Synonym: mitogen-activated protein kinase 8 isoform jnk1 beta2, mitogen-activated protein kinase 8, isoform cra_d, cdna flj77387, highly similar to homo sapiens mitogen-activated protein kinase 8 (mapk8), transcript variant 4, mRNA. Engineered: yes. Mutation: yes.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Organism_taxid: 9606

Resolution:

3.24Å R-factor: 0.195 R-free: 0.284

Authors:

T.Nakaniwa,T.Kinoshita,T.Inoue

Key ref:

T.Nakaniwa et al. (2012). Seven cysteine-deficient mutants depict the interplay between thermal and chemical stabilities of individual cysteine residues in mitogen-activated protein kinase c-Jun N-terminal kinase 1. Biochemistry, 51, 8410-8421. PubMed id: 23020677

Date:

28-Jun-12 Release date: 13-Feb-13

Protein chain

P45983  (MK08_HUMAN) -  Mitogen-activated protein kinase 8 from Homo sapiens

Seq: 427 a.a.

Struc: 356 a.a.*

* PDB and UniProt seqs differ at 11 residue positions (black crosses)

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

Biochemistry 51:8410-8421 (2012)

PubMed id: 23020677

Seven cysteine-deficient mutants depict the interplay between thermal and chemical stabilities of individual cysteine residues in mitogen-activated protein kinase c-Jun N-terminal kinase 1.

T.Nakaniwa, H.Fukada, T.Inoue, M.Gouda, R.Nakai, Y.Kirii, M.Adachi, T.Tamada, S.Segawa, R.Kuroki, T.Tada, T.Kinoshita.

ABSTRACT

Intracellular proteins can have free cysteines that may contribute to their structure, function, and stability; however, free cysteines can lead to chemical instabilities in solution because of oxidation-driven aggregation. The MAP kinase, c-Jun N-terminal kinase 1 (JNK1), possesses seven free cysteines and is an important drug target for autoimmune diseases, cancers, and apoptosis-related diseases. To characterize the role of cysteine residues in the structure, function, and stability of JNK1, we prepared and evaluated wild-type JNK1 and seven cysteine-deficient JNK1 proteins. The nonreduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis experiments showed that the chemical stability of JNK1 increased as the number of cysteines decreased. The contribution of each cysteine residue to biological function and thermal stability was highly susceptible to the environment surrounding the particular cysteine mutation. The mutations of solvent-exposed cysteine to serine did not influence biological function and increased the thermal stability. The mutation of the accessible cysteine involved in the hydrophobic pocket did not affect biological function, although a moderate thermal destabilization was observed. Cysteines in the loosely assembled hydrophobic environment moderately contributed to thermal stability, and the mutations of these cysteines had a negligible effect on enzyme activity. The other cysteines are involved in the tightly filled hydrophobic core, and mutation of these residues was found to correlate with thermal stability and enzyme activity. These findings about the role of cysteine residues should allow us to obtain a stable JNK1 and thus promote the discovery of potent JNK1 inhibitors.