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Literature summary for 2.7.7.67 extracted from

  • Ren, S.; Caforio, A.; Yang, Q.; Sun, B.; Yu, F.; Zhu, X.; Wang, J.; Dou, C.; Fu, Q.; Huang, N.; Sun, Q.; Nie, C.; Qi, S.; Gong, X.; He, J.; Wei, Y.; Driessen, A.J.; Cheng, W.
    Structural and mechanistic insights into the biosynthesis of CDP-archaeol in membranes (2017), Cell Res., 27, 1378-1391 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.67

Cloned(Commentary)

Cloned (Comment) Organism
gene carS, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) Aeropyrum pernix

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant enzyme in the CTP- and Mg2+-bound state, mixing of 25 mg/ml protein in 10 mM Tris-HCl, pH 8.0, containing 100 mM NaCl, 0.02% dodecyl-beta-D-maltopyranoside, and 10% glycerol with 200 mM NaCl, 20% w/v PEG 400, and 100 mM Tris-HCl, pH 8.0, at 20°C, 3 weeks, X-ray diffraction structure determination and analysis at 2.4 A resolution, structure modeling Aeropyrum pernix

Protein Variants

Protein Variants Comment Organism
A29W site-directed mutagenesis, the mutant shows no 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity Aeropyrum pernix
D100A site-directed mutagenesis, the mutant shows weak 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity compared to wild-type Aeropyrum pernix
D122A site-directed mutagenesis, the mutant shows no 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity Aeropyrum pernix
D125A site-directed mutagenesis, the mutant shows no 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity Aeropyrum pernix
F126A site-directed mutagenesis, the mutant shows no 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity Aeropyrum pernix
H157A site-directed mutagenesis, the mutant exhibits enhanced DGGGP-binding activity compared to wild-type Aeropyrum pernix
K57A site-directed mutagenesis, the mutant shows weak 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity compared to wild-type Aeropyrum pernix
L124W site-directed mutagenesis, the mutant shows no 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity Aeropyrum pernix
L156W site-directed mutagenesis, the mutant shows no 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity Aeropyrum pernix
N28A site-directed mutagenesis, the mutant shows weak 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate (DGGGP) substrate binding activity compared to wild-type Aeropyrum pernix
Q123A site-directed mutagenesis, the mutant exhibits enhanced DGGGP-binding activity compared to wild-type Aeropyrum pernix

Inhibitors

Inhibitors Comment Organism Structure
EDTA the inhibition is rescued by addition of of Mg2+ or Mn2+ but not Ca2+ or Zn2+ Aeropyrum pernix

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0002
-
 CTP in presence of Mg2+, pH 7.5, 37°C Aeropyrum pernix
0.00167
-
 CTP in absence of Mg2+, pH 7.5, 37°C Aeropyrum pernix

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane the enzyme comprises a transmembrane (TM) domain with five helices and cytoplasmic loops that together form a large charged cavity. The cytoplasmic cavity primarily comprises TM1, TM2, TM3 and TM4, and is loosely occluded by TM5. The remaining portion of the cavity is formed by two cytoplasmic loops (CLs): CL1 (between TM1 and TM2) and CL2 (between TM3 and TM4). CL1 and CL2 form the cytoplasmic domain (CPD), which caps the transmembrance domain (TMD). CL1, which is longer than CL2, is stabilized by packing against one side of TM3 Aeropyrum pernix 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
K+ activates Aeropyrum pernix
Li+ activates Aeropyrum pernix
Mg2+ required, activates Aeropyrum pernix
Mn2+ activates Aeropyrum pernix
additional information the Mg2+-dependent activity of ApCarS is enhanced by the addition of K+ or Li+ but not Na+ Aeropyrum pernix

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate Aeropyrum pernix
-
 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate Aeropyrum pernix ATCC 700893
-
 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate Aeropyrum pernix DSM 11879
-
 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate Aeropyrum pernix JCM 9820
-
 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate Aeropyrum pernix NBRC 100138
-
 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?

Organism

Organism UniProt Comment Textmining
Aeropyrum pernix Q9YF05
-
-
Aeropyrum pernix ATCC 700893 Q9YF05
-
-
Aeropyrum pernix DSM 11879 Q9YF05
-
-
Aeropyrum pernix JCM 9820 Q9YF05
-
-
Aeropyrum pernix NBRC 100138 Q9YF05
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Aeropyrum pernix

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate
-
 Aeropyrum pernix diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate ApCarS is a CTP transferase Aeropyrum pernix diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate
-
 Aeropyrum pernix ATCC 700893 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate ApCarS is a CTP transferase Aeropyrum pernix ATCC 700893 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate
-
 Aeropyrum pernix DSM 11879 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate ApCarS is a CTP transferase Aeropyrum pernix DSM 11879 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate
-
 Aeropyrum pernix JCM 9820 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate ApCarS is a CTP transferase Aeropyrum pernix JCM 9820 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate
-
 Aeropyrum pernix NBRC 100138 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
CTP + 2,3-bis-(O-geranylgeranyl)-sn-glycerol 1-phosphate ApCarS is a CTP transferase Aeropyrum pernix NBRC 100138 diphosphate + CDP-2,3-bis-(O-geranylgeranyl)-sn-glycerol
-
 ?
additional information purified ApCarS exhibits strong CTP-binding activity, binding site structure, overview Aeropyrum pernix ?
-
-
additional information purified ApCarS exhibits strong CTP-binding activity, binding site structure, overview Aeropyrum pernix ATCC 700893 ?
-
-
additional information purified ApCarS exhibits strong CTP-binding activity, binding site structure, overview Aeropyrum pernix DSM 11879 ?
-
-
additional information purified ApCarS exhibits strong CTP-binding activity, binding site structure, overview Aeropyrum pernix JCM 9820 ?
-
-
additional information purified ApCarS exhibits strong CTP-binding activity, binding site structure, overview Aeropyrum pernix NBRC 100138 ?
-
-

Synonyms

Synonyms Comment Organism
ApCarS
-
 Aeropyrum pernix
CarS
-
 Aeropyrum pernix
CDP-archaeol synthase
-
 Aeropyrum pernix

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
90
-
-
 Aeropyrum pernix

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
25 90 assays at Aeropyrum pernix

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Aeropyrum pernix

General Information

General Information Comment Organism
evolution structural comparisons and analyses, combined with functional studies, not only elucidated the mechanism governing the biosynthesis of phospholipids with ether-bonded isoprenoid chains by CTP transferase, but also provided insights into the evolution of this enzyme superfamily from archaea to bacteria and eukaryotes Aeropyrum pernix
additional information the enzyme comprises a transmembrane domain with five helices and cytoplasmic loops that together form a large charged cavity providing a binding site for CTP, modeling of the specific lipophilic substrate-binding site. Archaeol binds within two hydrophobic membrane-embedded grooves formed by the flexible transmembrane helix 5 (TM5), together with TM1 and TM4. CTP binds to one side of the central cavity of ApCarS and stabilizes the CPD through tight contacts. Recognition of CTP by ApCarS occurs through a combination of extensive polar and hydrophobic interactions, detailed overview Aeropyrum pernix
physiological function ApCarS is a CTP transferase Aeropyrum pernix