PDBsum entry 5dxf

Hydrolase

PDB id: 5dxf

Contents

Protein chains

460 a.a.

Waters ×98

PDB id:

5dxf

Name:

Hydrolase

Title:

Structure of candida albicans trehalose-6-phosphate phosphatase n- terminal domain

Structure:

Trehalose-6-phosphate phosphatase. Chain: a, b. Fragment: n-terminal domain (unp residues 1-534). Synonym: trehalose-phosphatase. Engineered: yes

Source:

Candida albicans (strain sc5314 / atcc mya- 2876). Yeast. Organism_taxid: 237561. Strain: sc5314 / atcc mya-2876. Gene: tps2, cao19.10556, cao19.3038, i503_00327, w5q_00328. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

2.56Å R-factor: 0.267 R-free: 0.298

Authors:

Y.Miao,R.G.Brennan

Key ref:

Y.Miao et al. (2016). Structures of trehalose-6-phosphate phosphatase from pathogenic fungi reveal the mechanisms of substrate recognition and catalysis. Proc Natl Acad Sci U S A, 113, 7148-7153. PubMed id: 27307435 DOI: 10.1073/pnas.1601774113

Date:

23-Sep-15 Release date: 15-Jun-16

Protein chains

Q5AI14  (Q5AI14_CANAL) -  Trehalose-phosphatase from Candida albicans (strain SC5314 / ATCC MYA-2876)

Seq: 888 a.a.

Struc: 460 a.a.*

* PDB and UniProt seqs differ at 5 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.4.1.15 - alpha,alpha-trehalose-phosphate synthase (UDP-forming).
• Reaction: D-glucose 6-phosphate + UDP-alpha-D-glucose = alpha,alpha-trehalose 6-phosphate + UDP + H⁺

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

reference

DOI no: 10.1073/pnas.1601774113

Proc Natl Acad Sci U S A 113:7148-7153 (2016)

PubMed id: 27307435

Structures of trehalose-6-phosphate phosphatase from pathogenic fungi reveal the mechanisms of substrate recognition and catalysis.

Y.Miao, J.L.Tenor, D.L.Toffaletti, E.J.Washington, J.Liu, W.R.Shadrick, M.A.Schumacher, R.E.Lee, J.R.Perfect, R.G.Brennan.

ABSTRACT

Trehalose is a disaccharide essential for the survival and virulence of pathogenic fungi. The biosynthesis of trehalose requires trehalose-6-phosphate synthase, Tps1, and trehalose-6-phosphate phosphatase, Tps2. Here, we report the structures of the N-terminal domain of Tps2 (Tps2NTD) from Candida albicans, a transition-state complex of the Tps2 C-terminal trehalose-6-phosphate phosphatase domain (Tps2PD) bound to BeF3 and trehalose, and catalytically dead Tps2PD(D24N) from Cryptococcus neoformans bound to trehalose-6-phosphate (T6P). The Tps2NTD closely resembles the structure of Tps1 but lacks any catalytic activity. The Tps2PD-BeF3-trehalose and Tps2PD(D24N)-T6P complex structures reveal a "closed" conformation that is effected by extensive interactions between each trehalose hydroxyl group and residues of the cap and core domains of the protein, thereby providing exquisite substrate specificity. Disruption of any of the direct substrate-protein residue interactions leads to significant or complete loss of phosphatase activity. Notably, the Tps2PD-BeF3-trehalose complex structure captures an aspartyl-BeF3 covalent adduct, which closely mimics the proposed aspartyl-phosphate intermediate of the phosphatase catalytic cycle. Structures of substrate-free Tps2PD reveal an "open" conformation whereby the cap and core domains separate and visualize the striking conformational changes effected by substrate binding and product release and the role of two hinge regions centered at approximately residues 102-103 and 184-188. Significantly, tps2Δ, tps2NTDΔ, and tps2D705N strains are unable to grow at elevated temperatures. Combined, these studies provide a deeper understanding of the substrate recognition and catalytic mechanism of Tps2 and provide a structural basis for the future design of novel antifungal compounds against a target found in three major fungal pathogens.