PDBsum entry 3t2c

Lyase, hydrolase

PDB id: 3t2c

Contents

Protein chain

389 a.a.

Ligands

13P ×2

Metals

_MG ×3

Waters ×373

PDB id:

3t2c

Name:

Lyase, hydrolase

Title:

Fructose-1,6-bisphosphate aldolase/phosphatase from thermoproteus neutrophilus, dhap-bound form

Structure:

Fructose-1,6-bisphosphate aldolase/phosphatase. Chain: a. Engineered: yes

Source:

Thermoproteus neutrophilus. Organism_taxid: 444157. Strain: dsm 2338 / jcm 9278 / v24sta. Gene: tneu_0133. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.30Å R-factor: 0.109 R-free: 0.129

Authors:

J.Du,R.Say,W.Lue,G.Fuchs,O.Einsle

Key ref:

J.Du et al. (2011). Active-site remodelling in the bifunctional fructose-1,6-bisphosphate aldolase/phosphatase. Nature, 478, 534-537. PubMed id: 21983965

Date:

22-Jul-11 Release date: 26-Oct-11

Protein chain

B1YAL1  (FBPAP_PYRNV) -  Fructose-1,6-bisphosphate aldolase/phosphatase from Pyrobaculum neutrophilum (strain DSM 2338 / JCM 9278 / NBRC 100436 / V24Sta)

Seq: 399 a.a.

Struc: 389 a.a.

Enzyme reactions

• Enzyme class 1: E.C.3.1.3.11 - fructose-bisphosphatase.
• Pathway: Pentose Phosphate Pathway (later stages)
• Reaction: beta-D-fructose 1,6-bisphosphate + H2O = beta-D-fructose 6-phosphate + phosphate

beta-D-fructose 1,6-bisphosphate + H2O = beta-D-fructose 6-phosphate (Bound ligand (Het Group name = 13P) matches with 62.50% similarity) + phosphate

• Enzyme class 2: E.C.4.1.2.13 - fructose-bisphosphate aldolase.
• Reaction: beta-D-fructose 1,6-bisphosphate = D-glyceraldehyde 3-phosphate + dihydroxyacetone phosphate

beta-D-fructose 1,6-bisphosphate = D-glyceraldehyde 3-phosphate (Bound ligand (Het Group name = 13P) corresponds exactly) + dihydroxyacetone phosphate

• Cofactor: Zn(2+)

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

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

reference

Nature 478:534-537 (2011)

PubMed id: 21983965

Active-site remodelling in the bifunctional fructose-1,6-bisphosphate aldolase/phosphatase.

J.Du, R.F.Say, W.Lü, G.Fuchs, O.Einsle.

ABSTRACT

Fructose-1,6-bisphosphate (FBP) aldolase/phosphatase is a bifunctional, thermostable enzyme that catalyses two subsequent steps in gluconeogenesis in most archaea and in deeply branching bacterial lineages. It mediates the aldol condensation of heat-labile dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (GAP) to FBP, as well as the subsequent, irreversible hydrolysis of the product to yield the stable fructose-6-phosphate (F6P) and inorganic phosphate; no reaction intermediates are released. Here we present a series of structural snapshots of the reaction that reveal a substantial remodelling of the active site through the movement of loop regions that create different catalytic functionalities at the same location. We have solved the three-dimensional structures of FBP aldolase/phosphatase from thermophilic Thermoproteus neutrophilus in a ligand-free state as well as in complex with the substrates DHAP and FBP and the product F6P to resolutions up to 1.3 Å. In conjunction with mutagenesis data, this pinpoints the residues required for the two reaction steps and shows that the sequential binding of additional Mg(2+) cations reversibly facilitates the reaction. FBP aldolase/phosphatase is an ancestral gluconeogenic enzyme optimized for high ambient temperatures, and our work resolves how consecutive structural rearrangements reorganize the catalytic centre of the protein to carry out two canonical reactions in a very non-canonical type of bifunctionality.