PDBsum entry 5d9h

Transferase

PDB id: 5d9h

Contents

Protein chains

290 a.a.

Ligands

ATP ×2

SO4 ×5

Metals

_MG ×2

PDB id:

5d9h

Name:

Transferase

Title:

Crystal structure of spak (stk39) dimer in the basal activity state

Structure:

Ste20/sps1-related proline-alanine-rich protein kinase. Chain: a, b. Fragment: unp residues 63-403. Synonym: ste-20-related kinase,serine/threonine-protein kinase 39. Engineered: yes

Source:

Mus musculus. Mouse. Organism_taxid: 10090. Gene: stk39, spak. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

3.10Å R-factor: 0.240 R-free: 0.269

Authors:

C.A.Taylor,Y.C.Juang,E.J.Goldsmith,M.H.Cobb

Key ref:

C.A.Taylor et al. (2015). Domain-Swapping Switch Point in Ste20 Protein Kinase SPAK. Biochemistry, 54, 5063-5071. PubMed id: 26208601 DOI: 10.1021/acs.biochem.5b00593

Date:

18-Aug-15 Release date: 02-Sep-15

Protein chains

Q9Z1W9  (STK39_MOUSE) -  STE20/SPS1-related proline-alanine-rich protein kinase from Mus musculus

Seq: 556 a.a.

Struc: 290 a.a.

Enzyme reactions

• Enzyme class: E.C.2.7.11.1 - non-specific serine/threonine 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⁺

L-seryl-[protein] (Bound ligand (Het Group name = ATP) corresponds exactly) + ATP = O-phospho-L-seryl-[protein] + ADP + H(+)

L-threonyl-[protein] (Bound ligand (Het Group name = ATP) corresponds exactly) + ATP = O-phospho-L-threonyl-[protein] + ADP + H(+)

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

reference

DOI no: 10.1021/acs.biochem.5b00593

Biochemistry 54:5063-5071 (2015)

PubMed id: 26208601

Domain-Swapping Switch Point in Ste20 Protein Kinase SPAK.

C.A.Taylor, Y.C.Juang, S.Earnest, S.Sengupta, E.J.Goldsmith, M.H.Cobb.

ABSTRACT

The related protein kinases SPAK and OSR1 regulate ion homeostasis in part by phosphorylating cation cotransporter family members. The structure of the kinase domain of OSR1 was determined in the unphosphorylated inactive form and, like some other Ste20 kinases, exhibited a domain-swapped activation loop. To further probe the role of domain swapping in SPAK and OSR1, we have determined the crystal structures of SPAK 63-403 at 3.1 Å and SPAK 63-390 T243D at 2.5 Å resolution. These structures encompass the kinase domain and different portions of the C-terminal tail, the longer without and the shorter with an activating T243D point mutation. The structure of the T243D protein reveals significant conformational differences relative to unphosphorylated SPAK and OSR1 but also has some features of an inactive kinase. Both structures are domain-swapped dimers. Sequences involved in domain swapping were identified and mutated to create a SPAK monomeric mutant with kinase activity, indicating that monomeric forms are active. The monomeric mutant is activated by WNK1 but has reduced activity toward its substrate NKCC2, suggesting regulatory roles for domain swapping. The structure of partially active SPAK T243D is consistent with a multistage activation process in which phosphorylation induces a SPAK conformation that requires further remodeling to build the active structure.