PDBsum entry 4z1y

Lyase

PDB id: 4z1y

Contents

Protein chains

424 a.a.

Ligands

2PG ×2

Metals

_MG ×2

Waters ×23

PDB id:

4z1y

Name:

Lyase

Title:

Thermostable enolase from chloroflexus aurantiacus with substrate 2- phosphoglycerate

Structure:

Enolase. Chain: a, b. Synonym: 2-phospho-d-glycerate hydro-lyase,2-phosphoglycerate dehydratase. Ec: 4.2.1.11

Source:

Chloroflexus aurantiacus (strain atcc 29366 / dsm 635 / j-10-fl). Organism_taxid: 324602. Strain: atcc 29366 / dsm 635 / j-10-fl. Atcc: 29366

Resolution:

2.53Å R-factor: 0.216 R-free: 0.252

Authors:

O.A.Zadvornyy,J.W.Peters

Key ref:

O.A.Zadvornyy et al. (2015). Biochemical and Structural Characterization of Enolase from Chloroflexus aurantiacus: Evidence for a Thermophilic Origin. Front Bioeng Biotechnol, 3, 74. PubMed id: 26082925 DOI: 10.3389/fbioe.2015.00074

Date:

27-Mar-15 Release date: 01-Jul-15

Protein chains

A9WCM4  (ENO_CHLAA) -  Enolase from Chloroflexus aurantiacus (strain ATCC 29366 / DSM 635 / J-10-fl)

Seq: 426 a.a.

Struc: 424 a.a.

Enzyme reactions

• Enzyme class: E.C.4.2.1.11 - phosphopyruvate hydratase.
• Reaction: (2R)-2-phosphoglycerate = phosphoenolpyruvate + H2O

(2R)-2-phosphoglycerate = phosphoenolpyruvate + H2O (Bound ligand (Het Group name = 2PG) matches with 90.91% similarity)

• Cofactor: Mg(2+)

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

Key reference

DOI no: 10.3389/fbioe.2015.00074

Front Bioeng Biotechnol 3:74 (2015)

PubMed id: 26082925

Biochemical and Structural Characterization of Enolase from Chloroflexus aurantiacus: Evidence for a Thermophilic Origin.

O.A.Zadvornyy, E.S.Boyd, M.C.Posewitz, N.A.Zorin, J.W.Peters.

ABSTRACT

Enolase catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate during both glycolysis and gluconeogenesis, and is required by all three domains of life. Here, we report the purification and biochemical and structural characterization of enolase from Chloroflexus aurantiacus, a thermophilic anoxygenic phototroph affiliated with the green non-sulfur bacteria. The protein was purified as a homodimer with a subunit molecular weight of 46 kDa. The temperature optimum for enolase catalysis was 80°C, close to the measured thermal stability of the protein which was determined to be 75°C, while the pH optimum for enzyme activity was 6.5. The specific activities of purified enolase determined at 25 and 80°C were 147 and 300 U mg(-1) of protein, respectively. K m values for the 2-phosphoglycerate/phosphoenolpyruvate reaction determined at 25 and 80°C were 0.16 and 0.03 mM, respectively. The K m values for Mg(2+) binding at these temperatures were 2.5 and 1.9 mM, respectively. When compared to enolase from mesophiles, the biochemical and structural properties of enolase from C. aurantiacus are consistent with this being thermally adapted. These data are consistent with the results of our phylogenetic analysis of enolase, which reveal that enolase has a thermophilic origin.