PDBsum entry: 2jog

Hydrolase

PDB id: 2jog

Contents

Protein chains

327 a.a. *

16 a.a. *

* Residue conservation analysis

PDB id:

2jog

Name:

Hydrolase

Title:

Structure of the calcineurin-nfat complex

Structure:

Calmodulin-dependent calcineurin a subunit alpha isoform. Chain: a. Fragment: residues 21-347. Synonym: serine/threonine-protein phosphatase 2b catalytic subunit alpha isoform, cam-prp catalytic subunit. Engineered: yes. Nfat. Chain: b.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: ppp3ca, calna, cna. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Other_details: the author states that this is an artificial sequence based on the nfat sequences. The

NMR struc:

10 models

Authors:

K.Takeuchi,M.H.Roehrl,Z.Y.Sun,G.Wagner

Key ref:

K.Takeuchi et al. (2007). Structure of the Calcineurin-NFAT Complex: Defining a T Cell Activation Switch Using Solution NMR and Crystal Coordinates. Structure, 15, 587-597. PubMed id: 17502104 DOI: 10.1016/j.str.2007.03.015

Date:

10-Mar-07 Release date: 22-May-07

Protein chain

Q08209  (PP2BA_HUMAN) -  Serine/threonine-protein phosphatase 2B catalytic subunit alpha isoform

Seq: 521 a.a.

Struc: 327 a.a.

Protein chain

No UniProt id for this chain

Enzyme reactions

• Enzyme class: Chain A: E.C.3.1.3.16 - Phosphoprotein phosphatase.
• Reaction: A phosphoprotein + H₂O = a protein + phosphate

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

 Gene Ontology (GO) functional annotation 

• Biochemical function: hydrolase activity (1 term)

Added reference

DOI no: 10.1016/j.str.2007.03.015

Structure 15:587-597 (2007)

PubMed id: 17502104

Structure of the Calcineurin-NFAT Complex: Defining a T Cell Activation Switch Using Solution NMR and Crystal Coordinates.

K.Takeuchi, M.H.Roehrl, Z.Y.Sun, G.Wagner.

ABSTRACT

Calcineurin (Cn) is a serine/threonine protein phosphatase that plays pivotal roles in many physiological processes, including cell proliferation, development, and apoptosis. Most prominently, Cn targets the nuclear factors of activated T cell (NFATs), transcription factors that activate cytokine genes. Calcium-activated Cn dephosphorylates multiple residues within the regulatory domain of NFAT, triggering joint nuclear translocation. This relies crucially on the interaction between the catalytic domain of Cn (CnCat) and the conserved PxIxIT motif located in a region distinct from the dephosphorylation sites of NFAT. Here, we present the structure of the complex between the 39 kDa CnCat and a 14 residue peptide containing a PVIVIT segment that was derived from affinity-driven peptide selection based on the conserved PxIxIT motif of NFATs. The structure of the complex was determined by using NMR assignments and structural constraints and the coordinates of the CnCat crystal structure. The NMR analysis relied on recently developed labeling and spectroscopic techniques. The VIVIT peptide is accommodated in a hydrophobic cleft formed by beta strands 11 and 14, and the loop between beta strands 11 and 12, forming a short parallel beta sheet with the exposed beta strand 14 in Cn. The side chains of conserved residues in the PxIxIT sequences make extensive interactions with conserved residues in Cn, while those of nonconserved residues are solvent exposed. The architecture of the interface explains the diversity of recognition sequences compatible with NFAT function and uncovers a potential targeting site for immune-suppressive agents. The structure reveals that the orientation of the bound PxIxIT directs the phosphorylation sites in NFAT's regulatory domain toward the Cn catalytic site.

Selected figure(s)

Figure 1.
Figure 1. Schematic Representation of the Domain Structure of Human Calcineurin and Human NFAT2
(A) For CnA, the locations of the binding sites for CnB, calmodulin (CaM), and the autoinhibitory domain (AI) are indicated with gray boxes. For CnB, four EF hand motifs are indicated.
(B) The location of the calcineurin binding site, the serine-rich region 1 (SRR1), the SPxx-repeat motifs 2 and 3 (SP2 and SP3), and nuclear localization and export signals of hNFAT2 are indicated.

Figure 6.
Figure 6. Interface of the CnCat-VIVIT Peptide Interaction
(A) Sites that are perturbed by the binding of the VIVIT peptide are mapped on the CnCat surface in the CnCat-VIVIT peptide complex. Red surfaces indicate sites that experienced both cross-saturation and chemical-shift perturbation. Yellow indicates the sites that showed chemical-shift changes upon binding of the VIVIT peptide. Blue residues were not affected in both experiments. White surfaces were not assigned. The VIVIT peptide is shown as a ribbon representation with the side chains for the PxIxIT motif.
(B) and (C) show the interface of CnCat and the conserved PxIxIT motif. In (B), the conserved residues in CnCat are shown with a colored surface representation based on the properties of each amino acid. Hydrophobic, acidic, basic, and hydrophilic residues are shown in yellow, red, blue, and white, respectively. The VIVIT peptide is shown as a ribbon representation with stick representation for the side-chain heavy atoms of the PxIxIT motif. The conserved Pro4, Ile6, Ile8, Thr9 are shown in dark red, while nonconserved Val6 and Val8 are colored in pink. In (C), the conserved residues in CnCat are shown as a stick model with color representation based on the properties of each amino acid. Hydrophobic, acidic, basic, and hydrophilic residues are shown in yellow, red, blue, and light blue, respectively. VIVIT is shown in the same representation as in (B).
(D) Schematic representation for the interaction between the PxIxI sequence and hydrophobic pockets of CnCat. The VIVIT peptide binds on the hydrophobic floor formed by Phe195, Tyr324, Val328, Met329, and Ile331. The conserved PxIxI sequence is accommodated in the hydrophobic pockets separated by Leu275, Tyr288, Met290, and Phe299. The peptide backbone is represented by the red arrow, and conserved and nonconserved side chains are shown in dark and light red, respectively.

The above figures are reprinted from an Open Access publication published by Cell Press: Structure (2007, 15, 587-597) copyright 2007.

Figures were selected by the author.