PMID- 10085115
OWN - NLM
STAT- MEDLINE
DCOM- 19990429
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 13
DP  - 1999 Mar 26
TI  - Effects of phosphorylation of threonine 160 on cyclin-dependent kinase 2
      structure and activity.
PG  - 8746-56
AB  - 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.
FAU - Brown, N R
AU  - Brown NR
AD  - Laboratory of Molecular Biophysics, Department of Biochemistry, and Oxford Centre
      for Molecular Sciences, University of Oxford, The Rex Richards Building, South
      Parks Road, Oxford OX1 3QU, United Kingdom.
FAU - Noble, M E
AU  - Noble ME
FAU - Lawrie, A M
AU  - Lawrie AM
FAU - Morris, M C
AU  - Morris MC
FAU - Tunnah, P
AU  - Tunnah P
FAU - Divita, G
AU  - Divita G
FAU - Johnson, L N
AU  - Johnson LN
FAU - Endicott, J A
AU  - Endicott JA
LA  - eng
SI  - PDB/1B38
SI  - PDB/1B39
SI  - PDB/R1B38SF
SI  - PDB/R1B39SF
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (Cyclin-Dependent Kinase Inhibitor Proteins)
RN  - 0 (Cyclins)
RN  - 0 (Histones)
RN  - 0 (Phosphoproteins)
RN  - 2ZD004190S (Threonine)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 2.7.- (Protein Kinases)
RN  - EC 2.7.1.- (histone H1 kinase)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.11.22 (CDC2-CDC28 Kinases)
RN  - EC 2.7.11.22 (CDK2 protein, human)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinase 2)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinases)
RN  - EC 2.7.11.22 (cyclin-dependent kinase-activating kinase)
RN  - EC 3.1.3.48 (CDKN3 protein, human)
RN  - EC 3.1.3.48 (Dual-Specificity Phosphatases)
RN  - EC 3.1.3.48 (Protein Tyrosine Phosphatases)
SB  - IM
MH  - Adenosine Triphosphate/metabolism
MH  - *CDC2-CDC28 Kinases
MH  - *Cell Cycle Proteins
MH  - Crystallography, X-Ray
MH  - Cyclin-Dependent Kinase 2
MH  - Cyclin-Dependent Kinase Inhibitor Proteins
MH  - Cyclin-Dependent Kinases/*chemistry/metabolism
MH  - Cyclins/metabolism
MH  - Dual-Specificity Phosphatases
MH  - Histones/metabolism
MH  - Humans
MH  - Kinetics
MH  - Models, Molecular
MH  - Phosphoproteins/chemistry
MH  - Phosphorylation
MH  - Protein Binding
MH  - Protein Conformation
MH  - Protein Kinases/metabolism
MH  - Protein Structure, Secondary
MH  - Protein Tyrosine Phosphatases/metabolism
MH  - Protein-Serine-Threonine Kinases/*chemistry/metabolism
MH  - Saccharomyces cerevisiae/enzymology
MH  - Threonine/*metabolism
EDAT- 1999/03/20 00:00
MHDA- 1999/03/20 00:01
CRDT- 1999/03/20 00:00
PHST- 1999/03/20 00:00 [pubmed]
PHST- 1999/03/20 00:01 [medline]
PHST- 1999/03/20 00:00 [entrez]
AID - 10.1074/jbc.274.13.8746 [doi]
AID - S0021-9258(19)87392-1 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Mar 26;274(13):8746-56. doi: 10.1074/jbc.274.13.8746.