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Literature summary extracted from

  • Montgomery, M.G.; Coker, A.R.; Taylor, I.A.; Wood, S.P.
    Assembly of a 20-nm protein cage by Escherichia coli 2-hydroxypentadienoic acid hydratase (2010), J. Mol. Biol., 396, 1379-1391.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.80 ligated into a modified pET22b expression vector encoding a His6 tag and expressed in Escherichia coli BL21(DE3) cells Escherichia coli K-12

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.2.1.80 MhpD with and without Mg2+, by hanging-drop vapour-diffusion method, to 2.8 A resolution. Space group of the crystals is H3 with unit cell parameters a = 207.35 A, b = 207.35 A, c = 545.47 A, alpha = 90°, beta = 90°, and gamma = 120°. Assembles to form a 20-nm-diameter particle comprising 60 protein subunits, arranged with 532 symmetry when crystallised at low pH in the presence of phosphate or sulphate ions. Particles form rapidly and are stable in solution during gel filtration at low pH. They are probably formed through trimers of pentamers, which are stabilised by the interaction of two phosphate ions with residues of the N-terminal domains of subunits at the 3fold axis. Once the particles are formed at high concentrations of phosphate (or sulphate), they remain stable in solution at 20fold lower concentrations of the anion. Guest molecules can be trapped within the hollow protein shell during assembly. The C-termini of the subunits are freely accessible on the surface of the protein cage Escherichia coli K-12

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.80 Mg2+ active site of MhpD consists of three acidic residues (Glu105, Glu107, and Glu138), which may be involved in coordinating the divalent cation essential for activity Escherichia coli K-12

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.80 30000
-
 5 * 35000, fusion protein, SDS-PAGE. 5 * 30000, purified protein, SDS-PAGE Escherichia coli K-12
4.2.1.80 35000
-
 5 * 35000, fusion protein, SDS-PAGE. 5 * 30000, purified protein, SDS-PAGE Escherichia coli K-12
4.2.1.80 151000
-
 gel filtration-multiangle laser light scattering Escherichia coli K-12

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.80 Escherichia coli K-12 P77608
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.80 by centrifugation and gel filtration Escherichia coli K-12

Subunits

EC Number Subunits Comment Organism
4.2.1.80 pentamer 5 * 35000, fusion protein, SDS-PAGE. 5 * 30000, purified protein, SDS-PAGE Escherichia coli K-12

Synonyms

EC Number Synonyms Comment Organism
4.2.1.80 2-hydroxypentadienoic acid hydratase
-
 Escherichia coli K-12
4.2.1.80 HPD hydratase
-
 Escherichia coli K-12
4.2.1.80 MhpD
-
 Escherichia coli K-12