PDBsum entry 2ozo

Transferase

PDB id: 2ozo

Contents

Protein chain

545 a.a. *

Ligands

ANP

Metals

_MG

Waters ×69

* Residue conservation analysis

PDB id:

2ozo

Name:

Transferase

Title:

Autoinhibited intact human zap-70

Structure:

Tyrosine-protein kinase zap-70. Chain: a. Fragment: inactive zap-70 (residues 1-606). Synonym: 70 kda zeta-associated protein, syk-related tyrosine kinase. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: zap70, srk. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108. Expression_system_cell: baculovirus-infected insect cells sf9.

Resolution:

2.60Å R-factor: 0.220 R-free: 0.290

Authors:

S.Deindl,T.A.Kadlecek,T.Brdicka,X.Cao,A.Weiss,J.Kuriyan

Key ref:

S.Deindl et al. (2007). Structural Basis for the Inhibition of Tyrosine Kinase Activity of ZAP-70. Cell, 129, 735-746. PubMed id: 17512407 DOI: 10.1016/j.cell.2007.03.039

Date:

26-Feb-07 Release date: 22-May-07

Protein chain

P43403  (ZAP70_HUMAN) -  Tyrosine-protein kinase ZAP-70 from Homo sapiens

Seq: 619 a.a.

Struc: 545 a.a.*

* PDB and UniProt seqs differ at 10 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.7.10.2 - non-specific protein-tyrosine kinase.
• Reaction: L-tyrosyl-[protein] + ATP = O-phospho-L-tyrosyl-[protein] + ADP + H⁺

L-tyrosyl-[protein] + ATP = O-phospho-L-tyrosyl-[protein] (Bound ligand (Het Group name = ANP) matches with 81.25% similarity) + ADP + H(+)

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

Added reference

DOI no: 10.1016/j.cell.2007.03.039

Cell 129:735-746 (2007)

PubMed id: 17512407

Structural Basis for the Inhibition of Tyrosine Kinase Activity of ZAP-70.

S.Deindl, T.A.Kadlecek, T.Brdicka, X.Cao, A.Weiss, J.Kuriyan.

ABSTRACT

ZAP-70, a cytoplasmic tyrosine kinase required for T cell antigen receptor signaling, is controlled by a regulatory segment that includes a tandem SH2 unit responsible for binding to immunoreceptor tyrosine-based activation motifs (ITAMs). The crystal structure of autoinhibited ZAP-70 reveals that the inactive kinase domain adopts a conformation similar to that of cyclin-dependent kinases and Src kinases. The autoinhibitory mechanism of ZAP-70 is, however, distinct and involves interactions between the regulatory segment and the hinge region of the kinase domain that reduce its flexibility. Two tyrosine residues in the SH2-kinase linker that activate ZAP-70 when phosphorylated are involved in aromatic-aromatic interactions that connect the linker to the kinase domain. These interactions are inconsistent with ITAM binding, suggesting that destabilization of this autoinhibited ZAP-70 conformation is the first step in kinase activation.

Selected figure(s)

Figure 5.
Figure 5. Mutagenesis Identifies Hot Spot Residues in the Linker-Kinase Sandwich
(A) Two schematic views of ZAP-70, rotated by 180° with respect to each other, depict the location of residues in ZAP-70 that are most critical for autoinhibition. These hot spot residues are indicated by yellow stars.
(B) LAT phosphorylation by ZAP-70 mutants. The Tyr315Ala/Tyr319Ala double mutant is referred to as “YYAA.” Strongly activating alanine mutations at hot spots of the linker-kinase sandwich are highlighted with yellow stars. Control experiments in the presence of Lck are shown in Figure S4.

Figure 6.
Figure 6. Interaction between the Inter-SH2 Linker and the Kinase Domain
Detailed view showing the docking of Pro147 in the inter-SH2 linker into the cleft formed by the side chains of Tyr597 and Tyr598. Helix αI is shown as a light brown surface, and Tyr597 and Tyr598 are shown in magenta surface representation.

The above figures are reprinted by permission from Cell Press: Cell (2007, 129, 735-746) copyright 2007.

Figures were selected by an automated process.