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Literature summary extracted from

  • Wang, H.; Pang, H.; Bartlam, M.; Rao, Z.
    Crystal Structure of Human E1 Enzyme and its Complex with a Substrate Analog Reveals the Mechanism of its Phosphatase/Enolase Activity (2005), J. Mol. Biol., 348, 917-926.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.3.77 into the bacterial expression vector pET28a with a His tag on the C-terminus. Recombinant plasmid overexpressed in Escherichia coli strain BL21 (DE3) Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.3.77 crystal structure to 1.7 A resolution of MASA and its complex with a substrate analog, by hanging-drop vapor-diffusion technique Homo sapiens
4.2.1.11 1.7 A resolution by multiwavelength anomalous diffraction and molecular replacement techniques Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.3.77 Mg2+ three magnesium ions, one is buried in the active-site pocket in the interface between the two domains, where it serves as a cofactor for the enzyme activity. The other two are located on the surface of the molecule and are supposedly helpful for packing of crystals. Magnesium ions originate from the reservoir solution during crystallization and are absolutely required for the enzyme stability and activity Homo sapiens
4.2.1.11 Mg2+
-
 Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.11 2,3-dioxo-5-methylthio-1-phosphopentane + 4 H+ Homo sapiens
-
 3-hydroxy-5-methyl-thio-pent-2-en-1-yl-phosphate + H2O
-
 ?
4.2.1.11 2-phospho-D-glycerate Homo sapiens
-
 phosphoenolpyruvate + H2O
-
 r

Organism

EC Number Organism UniProt Comment Textmining
3.1.3.77 Homo sapiens Q9UHY7
-
-
4.2.1.11 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.3.77 by Ni2+-chelating affinity column, gel filtration and anion-exchange chromatography, more than 90% pure Homo sapiens

Storage Stability

EC Number Storage Stability Organism
3.1.3.77 -80°C, 20 mM HEPES buffer, pH 7.5 Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.3.77 (1-oxo-heptyl)-phosphonic acid + H2O substrate analog, in complex with MASA it sits in the cleft formed by the conserved residues in the active site Homo sapiens ?
-
 ?
4.2.1.11 2,3-dioxo-5-methylthio-1-phosphopentane + 4 H+
-
 Homo sapiens 3-hydroxy-5-methyl-thio-pent-2-en-1-yl-phosphate + H2O
-
 ?
4.2.1.11 2-phospho-D-glycerate
-
 Homo sapiens phosphoenolpyruvate + H2O
-
 r

Subunits

EC Number Subunits Comment Organism
3.1.3.77 monomer
-
 Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
3.1.3.77 enolase-phosphatase
-
 Homo sapiens
3.1.3.77 MASA
-
 Homo sapiens
4.2.1.11 Enolase-phosphatase E1
-
 Homo sapiens
4.2.1.11 MASA
-
 Homo sapiens