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Literature summary extracted from

  • Liu, C.I.; Jeng, W.Y.; Chang, W.J.; Shih, M.F.; Ko, T.P.; Wang, A.H.
    Structural insights into the catalytic mechanism of human squalene synthase (2014), Acta Crystallogr. Sect. D, 70, 231-241.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.21 gene hSQS, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.21 purified recombinant wild-type and mutant enzymes in complex with Mg2+ and/or farnesyl diphosphate or presqualene diphosphate, respectively, mixing equal volumes of protein solution, containing 10 mg/ml protein and 3 mM substrate or 1 mM intermediate, with or without 1 mM Mg2+, with precipitant solution (form I: 20% PEG 2000 MME, 0.01 M NiCl2, 0.1 M Tris, pH 8.5 and form II: 1.4 M sodium citrate tribasic dehydrate, 0.1 M Na HEPES, pH 7.5; form III, 2 M K2HPO4/NaH2PO4, pH 6.5) at 25°C, X-ray diffraction structure determination and analysis at 1.75-2.35 A resolution, modelling Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.21 F288A site-directed mutagenesis, structure comparison with bound metals and reaction intermediate PSPP compared to the wild-type enzyme Homo sapiens
2.5.1.21 F288L site-directed mutagenesis, structure comparison with bound metals and reaction intermediate PSPP compared to the wild-type enzyme Homo sapiens
2.5.1.21 Y73A site-directed mutagenesis, structure comparison with bound metals and reaction intermediate PSPP compared to the wild-type enzyme Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.21 additional information
-
 additional information steady-state kinetics Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.21 Mg2+ the enzyme requires either Mg2+ or Mn2+ as metal cofactor Homo sapiens
2.5.1.21 Mn2+ the enzyme requires either Mg2+ or Mn2+ as metal cofactor Homo sapiens
2.5.1.21 additional information squalene synthase is a divalent metal-ion-dependent enzyme Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.21 2 (2E,6E)-farnesyl diphosphate Homo sapiens first half-reaction diphosphate + presqualene diphosphate
-
 ?
2.5.1.21 2 (2E,6E)-farnesyl diphosphate + NADPH + H+ Homo sapiens
-
 squalene + 2 diphosphate + NADP+
-
 ?
2.5.1.21 additional information Homo sapiens the catalytic process involves two reactions: firstly, two FPP molecules are condensed to form presqualene diphosphate (PSPP), an intermediate with a cyclopropane C1'-C2-C3 ring structure, and secondly, PSPP undergoes a NADPH-dependent rearrangement and reduction to generate the end product squalene ?
-
 ?
2.5.1.21 presqualene diphosphate + NADPH + H+ Homo sapiens second half reaction squalene + diphosphate + NADP+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.21 Homo sapiens P37268
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.21 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography, the N-terminal His-tag is then cleaved using thrombin and removed using nickel affinity chromatography Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.21 2 (2E,6E)-farnesyl diphosphate first half-reaction Homo sapiens diphosphate + presqualene diphosphate
-
 ?
2.5.1.21 2 (2E,6E)-farnesyl diphosphate + NADPH + H+
-
 Homo sapiens squalene + 2 diphosphate + NADP+
-
 ?
2.5.1.21 2 (2E,6E)-farnesyl diphosphate + NADPH + H+ overall reaction, the enzyme catalyzes the two-step reductive head-to-head condensation of two molecules of farnesyl pyrophosphate to form squalene using presqualene diphosphate (PSPP) as an intermediate Homo sapiens squalene + 2 diphosphate + NADP+
-
 ?
2.5.1.21 additional information the catalytic process involves two reactions: firstly, two FPP molecules are condensed to form presqualene diphosphate (PSPP), an intermediate with a cyclopropane C1'-C2-C3 ring structure, and secondly, PSPP undergoes a NADPH-dependent rearrangement and reduction to generate the end product squalene Homo sapiens ?
-
 ?
2.5.1.21 additional information catalytic mechanism, overview Homo sapiens ?
-
 ?
2.5.1.21 presqualene diphosphate + NADPH + H+ second half reaction Homo sapiens squalene + diphosphate + NADP+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.5.1.21 SQS
-
 Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
2.5.1.21 NADPH dependent on, NADPH-binding residues and modeling, overview Homo sapiens

General Information

EC Number General Information Comment Organism
2.5.1.21 metabolism four major steps - substrate binding, condensation, intermediate formation and translocation - of the ordered sequential mechanisms involved in the 1'-1 isoprenoid biosynthetic pathway Homo sapiens
2.5.1.21 additional information determination and analysis of human SQS and its mutants in complex with several substrate analogues and intermediates coordinated with Mg2+ or Mn2+, SQS active analysis, overview Homo sapiens