The structure of the cell cycle protein Cdc14 reveals a proline-directed protein phosphatase

被引:125
作者
Gray, CH
Good, VM
Tonks, NK
Barford, D
机构
[1] Inst Canc Res, Sect Struct Biol, Chester Beatty Labs, London SW3 6JB, England
[2] Cold Spring Harbor Lab, Cold Spring Harbor, NY 11724 USA
关键词
Cdc14; dual-specificity phosphatase; proline-directed phosphatase; protein structure; X-ray crystallography;
D O I
10.1093/emboj/cdg348
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The Cdc14 family of dual-specificity protein phosphatases (DSPs) is conserved within eukaryotes and functions to down-regulate mitotic Cdk activities, promoting cytokinesis and mitotic exit. We have integrated structural and kinetic analyses to define the molecular mechanism of the dephosphorylation reaction catalysed by Cdc14. The structure of Cdc14 illustrates a novel arrangement of two domains, each with a DSP-like fold, arranged in tandem. The C-terminal domain contains the conserved PTP motif of the catalytic site, whereas the N-terminal domain, which shares no sequence similarity with other DSPs, contributes to substrate specificity, and lacks catalytic activity. The catalytic site is located at the base of a pronounced surface channel formed by the interface of the two domains, and regions of both domains interact with the phosphopeptide substrate. Specificity for a pSer-Pro motif is mediated by a hydrophobic pocket that is capable of accommodating the apolar Pro(P+1) residue of the peptide. Our structural and kinetic data support a role for Cdc14 in the preferential dephosphorylation of proteins modified by proline-directed kinases.
引用
收藏
页码:3524 / 3535
页数:12
相关论文
共 41 条
[1]   Methods used in the structure determination of bovine mitochondrial F-1 ATPase [J].
Abrahams, JP ;
Leslie, AGW .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1996, 52 :30-42
[2]   THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY [J].
BAILEY, S .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 :760-763
[3]   The structure and mechanism of protein phosphatases: Insights into catalysis and regulation [J].
Barford, D ;
Das, AK ;
Egloff, MP .
ANNUAL REVIEW OF BIOPHYSICS AND BIOMOLECULAR STRUCTURE, 1998, 27 :133-164
[4]   Regulation of the anaphase-promoting complex by the dual specificity phosphatase human Cdc14a [J].
Bembenek, J ;
Yu, HT .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (51) :48237-48242
[5]   The Protein Data Bank [J].
Berman, HM ;
Westbrook, J ;
Feng, Z ;
Gilliland, G ;
Bhat, TN ;
Weissig, H ;
Shindyalov, IN ;
Bourne, PE .
NUCLEIC ACIDS RESEARCH, 2000, 28 (01) :235-242
[6]   The structural basis for specificity of substrate and recruitment peptides for cyclin-dependent kinases [J].
Brown, NR ;
Noble, MEM ;
Endicott, JA ;
Johnson, LN .
NATURE CELL BIOLOGY, 1999, 1 (07) :438-443
[7]   Crystallography & NMR system:: A new software suite for macromolecular structure determination [J].
Brunger, AT ;
Adams, PD ;
Clore, GM ;
DeLano, WL ;
Gros, P ;
Grosse-Kunstleve, RW ;
Jiang, JS ;
Kuszewski, J ;
Nilges, M ;
Pannu, NS ;
Read, RJ ;
Rice, LM ;
Simonson, T ;
Warren, GL .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1998, 54 :905-921
[8]   Structure and mechanism of the RNA triphosphatase component of mammalian mRNA capping enzyme [J].
Changela, A ;
Ho, CK ;
Martins, A ;
Shuman, S ;
Mondragón, A .
EMBO JOURNAL, 2001, 20 (10) :2575-2586
[9]  
Cueille N, 2001, J CELL SCI, V114, P2649
[10]   Maximum-likelihood heavy-atom parameter refinement for multiple isomorphous replacement and multiwavelength anomalous diffraction methods [J].
delaFortelle, E ;
Bricogne, G .
MACROMOLECULAR CRYSTALLOGRAPHY, PT A, 1997, 276 :472-494