PDBsum entry 4ouc

Transferase

PDB id: 4ouc

Contents

Protein chain

329 a.a.

Ligands

ALA-ARG-THR-LYS-GLN-THR-ALA

EDO ×6

5ID-IOD

Metals

_NA

Waters ×419

PDB id:

4ouc

Name:

Transferase

Title:

Structure of human haspin in complex with histone h3 substrate

Structure:

Serine/threonine-protein kinase haspin. Chain: a. Fragment: unp residues 465-798. Synonym: germ cell-specific gene 2 protein, h-haspin, haploid germ cell-specific nuclear protein kinase. Engineered: yes. Histone h3.2. Chain: b. Fragment: histone h3 tail, unp residues 2-13.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: gsg2. Expressed in: escherichia coli. Expression_system_taxid: 469008. Synthetic: yes. Organism_taxid: 9606

Resolution:

1.90Å R-factor: 0.166 R-free: 0.211

Authors:

A.Chaikuad,F.Von Delft,C.H.Arrowsmith,A.M.Edwards,C.Bountra,S.Knapp, Structural Genomics Consortium (Sgc)

Key ref:

A.Maiolica et al. (2014). Modulation of the chromatin phosphoproteome by the Haspin protein kinase. Mol Cell Proteomics, 13, 1724-1740. PubMed id: 24732914 DOI: 10.1074/mcp.M113.034819

Date:

15-Feb-14 Release date: 16-Apr-14

Protein chain

Q8TF76  (HASP_HUMAN) -  Serine/threonine-protein kinase haspin from Homo sapiens

Seq: 798 a.a.

Struc: 329 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] + ATP = O-phospho-L-seryl-[protein] (Bound ligand (Het Group name = 5ID) matches with 62.07% similarity) + ADP + H(+)

L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] (Bound ligand (Het Group name = 5ID) matches with 62.07% similarity) + ADP + H(+)

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

reference

DOI no: 10.1074/mcp.M113.034819

Mol Cell Proteomics 13:1724-1740 (2014)

PubMed id: 24732914

Modulation of the chromatin phosphoproteome by the Haspin protein kinase.

A.Maiolica, M.de Medina-Redondo, E.M.Schoof, A.Chaikuad, F.Villa, M.Gatti, S.Jeganathan, H.J.Lou, K.Novy, S.Hauri, U.H.Toprak, F.Herzog, P.Meraldi, L.Penengo, B.E.Turk, S.Knapp, R.Linding, R.Aebersold.

ABSTRACT

Recent discoveries have highlighted the importance of Haspin kinase activity for the correct positioning of the kinase Aurora B at the centromere. Haspin phosphorylates Thr(3) of the histone H3 (H3), which provides a signal for Aurora B to localize to the centromere of mitotic chromosomes. To date, histone H3 is the only confirmed Haspin substrate. We used a combination of biochemical, pharmacological, and mass spectrometric approaches to study the consequences of Haspin inhibition in mitotic cells. We quantified 3964 phosphorylation sites on chromatin-associated proteins and identified a Haspin protein-protein interaction network. We determined the Haspin consensus motif and the co-crystal structure of the kinase with the histone H3 tail. The structure revealed a unique bent substrate binding mode positioning the histone H3 residues Arg(2) and Lys(4) adjacent to the Haspin phosphorylated threonine into acidic binding pockets. This unique conformation of the kinase-substrate complex explains the reported modulation of Haspin activity by methylation of Lys(4) of the histone H3. In addition, the identification of the structural basis of substrate recognition and the amino acid sequence preferences of Haspin aided the identification of novel candidate Haspin substrates. In particular, we validated the phosphorylation of Ser(137) of the histone variant macroH2A as a target of Haspin kinase activity. MacroH2A Ser(137) resides in a basic stretch of about 40 amino acids that is required to stabilize extranucleosomal DNA, suggesting that phosphorylation of Ser(137) might regulate the interactions of macroH2A and DNA. Overall, our data suggest that Haspin activity affects the phosphorylation state of proteins involved in gene expression regulation and splicing.