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Literature summary extracted from

  • Kajikawa, M.; Kinohira, S.; Ando, A.; Shimoyama, M.; Kato, M.; Fukuzawa, H.
    Accumulation of squalene in a microalga Chlamydomonas reinhardtii by genetic modification of squalene synthase and squalene epoxidase genes (2015), PLoS ONE, 10, e0120446.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.21 gene CrSQS, a single-copy gene, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant functional expression of C-terminally His6-tagged enzyme, with the hydrophobic transmembrane regions removed, in Escherichia coli. Transient expression of full-length GFP-tagged CrSQS and GFP-tagged squalene epoxidase CrSQE in onion epidermal tissue suggests that both proteins are co-localized in the endoplasmic reticulum. Overexpression of gene CrSQS in Chlamydomonas reinhardtii mutant strain UVM4 under the regulation of the constitutive PsaD promoter, squalene does not accumulate significantly in CrSQS-overexpressing cells, although conversion of farnesyl diphosphate to squalene is enhanced by overexpression of enzyme CrSQS Chlamydomonas reinhardtii

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.21 additional information CrSQS-overexpression increases the rate of conversion of 14C-labeled farnesylpyrophosphate into squalene but does not lead to overaccumulation of squalene. Addition of terbinafine causes the accumulation of squalene and suppression of cell survival. In CrSQE-knockdown lines, the expression level of CrSQE is reduced by 59-76% of that in wild-type cells, and significant levels of squalene accumulate without any growth inhibition. In co-transformation lines with CrSQS-overexpression and squalene epoxidase CrSQE-knockdown, the level of squalene is not increased significantly compared with that in solitary CrSQE knockdown lines Chlamydomonas reinhardtii

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.5.1.21 endoplasmic reticulum membrane CrSQS protein has two putative transmembrane domains at the C-terminus, Phe398-Glu420 and Gln436-Leu458 Chlamydomonas reinhardtii 5789
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.21 Mg2+ the enzyme requires either Mg2+ or Mn2+ as metal cofactor Chlamydomonas reinhardtii
2.5.1.21 Mn2+ the enzyme requires either Mg2+ or Mn2+ as metal cofactor Chlamydomonas reinhardtii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.21 46000
-
-
 Chlamydomonas reinhardtii
2.5.1.21 57000
-
-
 Chlamydomonas reinhardtii

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 + NADPH + H+ Chlamydomonas reinhardtii
-
 squalene + 2 diphosphate + NADP+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.21 Chlamydomonas reinhardtii
-
 gene CrSQS
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.21 recombinant C-terminally His6-tagged and truncated enzyme from Escherichia coli by cobalt affinity chromatography Chlamydomonas reinhardtii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.21 2 (2E,6E)-farnesyl diphosphate + NADPH + H+
-
 Chlamydomonas reinhardtii squalene + 2 diphosphate + NADP+
-
 ?

Subunits

EC Number Subunits Comment Organism
2.5.1.21 ? x * 46000, recombinant C-terminally His6-tagged and truncated enzyme, SDS-PAGE, x * 57000, recombinant full-length enzyme, SDS-PAGE Chlamydomonas reinhardtii

Synonyms

EC Number Synonyms Comment Organism
2.5.1.21 CrSQS
-
 Chlamydomonas reinhardtii
2.5.1.21 SQS
-
 Chlamydomonas reinhardtii

Cofactor

EC Number Cofactor Comment Organism Structure
2.5.1.21 NADPH dependent on Chlamydomonas reinhardtii

General Information

EC Number General Information Comment Organism
2.5.1.21 metabolism the enzyme is involved in squalene synthesis and sterol metabolism Chlamydomonas reinhardtii
2.5.1.21 additional information squalene does not accumulate significantly in CrSQS-overexpressing cells, although conversion of farnesyl diphosphate to squalene is enhanced by overexpression of enzyme CrSQS Chlamydomonas reinhardtii
2.5.1.21 physiological function the enzyme is involved in squalene synthesis and sterol metabolism Chlamydomonas reinhardtii