PDBsum entry 1b39

Transferase

PDB id: 1b39

Contents

Protein chain

291 a.a. *

Ligands

ATP

Metals

_MG

Waters ×145

* Residue conservation analysis

PDB id:

1b39

Name:

Transferase

Title:

Human cyclin-dependent kinase 2 phosphorylated on thr 160

Structure:

Protein (cell division protein kinase 2). Chain: a. Fragment: intact. Synonym: p33 protein kinase. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108. Expression_system_cell_line: sf9.

Resolution:

2.10Å R-factor: 0.200 R-free: 0.270

Authors:

N.R.Brown,M.E.M.Noble,A.M.Lawrie,M.C.Morris,P.Tunnah,G.Divita, L.N.Johnson,J.A.Endicott

Key ref:

N.R.Brown et al. (1999). Effects of phosphorylation of threonine 160 on cyclin-dependent kinase 2 structure and activity. J Biol Chem, 274, 8746-8756. PubMed id: 10085115 DOI: 10.1074/jbc.274.13.8746

Date:

17-Dec-98 Release date: 23-Dec-98

Protein chain

P24941  (CDK2_HUMAN) -  Cyclin-dependent kinase 2 from Homo sapiens

Seq: 298 a.a.

Struc: 290 a.a.

Enzyme reactions

• Enzyme class: E.C.2.7.11.22 - cyclin-dependent 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: 10.1074/jbc.274.13.8746

J Biol Chem 274:8746-8756 (1999)

PubMed id: 10085115

Effects of phosphorylation of threonine 160 on cyclin-dependent kinase 2 structure and activity.

N.R.Brown, M.E.Noble, A.M.Lawrie, M.C.Morris, P.Tunnah, G.Divita, L.N.Johnson, J.A.Endicott.

ABSTRACT

We have prepared phosphorylated cyclin-dependent protein kinase 2 (CDK2) for crystallization using the CDK-activating kinase 1 (CAK1) from Saccharomyces cerevisiae and have grown crystals using microseeding techniques. Phosphorylation of monomeric human CDK2 by CAK1 is more efficient than phosphorylation of the binary CDK2-cyclin A complex. Phosphorylated CDK2 exhibits histone H1 kinase activity corresponding to approximately 0.3% of that observed with the fully activated phosphorylated CDK2-cyclin A complex. Fluorescence measurements have shown that Thr160 phosphorylation increases the affinity of CDK2 for both histone substrate and ATP and decreases its affinity for ADP. By contrast, phosphorylation of CDK2 has a negligible effect on the affinity for cyclin A. The crystal structures of the ATP-bound forms of phosphorylated CDK2 and unphosphorylated CDK2 have been solved at 2.1-A resolution. The structures are similar, with the major difference occurring in the activation segment, which is disordered in phosphorylated CDK2. The greater mobility of the activation segment in phosphorylated CDK2 and the absence of spontaneous crystallization suggest that phosphorylated CDK2 may adopt several different mobile states. The majority of these states are likely to correspond to inactive conformations, but a small fraction of phosphorylated CDK2 may be in an active conformation and hence explain the basal activity observed.

Selected figure(s)

Figure 2.
Fig. 2. CDK-associated phosphatase KAP dephosphorylates phosphorylated CDK2. KAP specifically dephosphorylates monomeric phosphorylated CDK2.

Figure 6.
Fig. 6. a, the fold of monomeric CDK2. The structure is shown in a schematic representation with regions of -sheet shown as arrows and -helix shown as ribbons. The N-terminal domain is colored principally white, with the exception of the glycine-rich loop (colored magenta), and the C-helix (PSTAIRE helix, colored gold). The region of the N-terminal domain for which no trace is visible (residues 36-43) is indicated by small black spheres identifying residues 35 and 44. ATP is shown in ball and stick representation at the interface between the N- and C-terminal domains. The C-terminal domain is colored pink, with the activation segment (residues 145-172) highlighted in cyan. b, comparison of electron density for the tip of the activation segment. The upper stereo pair shows electron density defining the conformation of residues at the tip of the activation segment (residues 155-165) in the ATP complex of unphosphorylated monomeric CDK2, while the lower stereo pair shows the equivalent electron density in phosphorylated monomeric CDK2. In this figure the phosphate group attached to Thr^160 has been omitted from the phosphorylated CDK2 structure for clarity. The maps were calculated using (2F[o] F[c]) [calc] coefficients generated by REFMAC and are contoured at a level of 0.2e^ Å^ 3. c, B-factor plots for CDK2-ATP and phosphorylated CDK2-ATP. The mean main chain B-factor of each residue along the polypeptide chain is shown for unphosphorylated CDK2 (thin lines) and phosphorylated CDK2 (thick lines). The outstanding regions of difference include the glycine loop (residues 8-18) and the tip of the activation segment (residues 155-165). d, detail of the fold of the CDK2-ATP complex. The interaction of Tyr^159 and Thr^160, at the tip of the activation segment, with residues Glu^12-Tyr^15 in the glycine-rich loop is shown. The coloring scheme is the same as for a.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (1999, 274, 8746-8756) copyright 1999.

Figures were selected by an automated process.