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Literature summary extracted from

  • Lu, X.; Lin, S.; Chang, C.C.Y.; Chang, T.
    Mutant acyl-coenzyme A: cholesterol acyltransferase 1 devoid of cysteine residues remains catalytically active (2002), J. Biol. Chem., 277, 711-718.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.26 expression of ACAT1 in infected insect High Five cells Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.26 C333A expressed in H5 cells, the mutant retains a significant amount of activity, Cys-333 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C333A/C345A/C365A expressed in H5 cells, the mutant contains essentially the same activity as the wild-type ACAT1. Sensitive to p-chloromercuribenzene sulfonic acid inhibition Homo sapiens
2.3.1.26 C345A expressed in H5 cells, the mutant retains a significant amount of activity, Cys-345 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C345A/C467A insensitive to p-chloromercuribenzene sulfonic acid Homo sapiens
2.3.1.26 C365A expressed in H5 cells, the mutant retains a significant amount of activity, Cys-365 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C387A expressed in H5 cells, the mutant retains a significant amount of activity, Cys-387 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C467A expressed in H5 cells, the mutant retains a significant amount of activity, Cys-467 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C516A expressed in H5 cells, the mutant retains a significant amount of activity, Cys-516 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C528A expressed in H5 cells, the mutant retains a significant amount of activity, Cys-528 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C546A expressed in H5 cells, the mutant retains a significant amount of activity , Cys-546 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C92A expressed in H5 cells, the mutant retains a significant amount of activity, Cys-92 is not essential for ACAT1 catalysis, the mutant is expressed as a single, undegraded 50-kDA band Homo sapiens
2.3.1.26 C92A/C333A/C345A/C365A/C387A/C467A/C516A resistant to p-chloromercuribenzene sulfonic acid inhibition Homo sapiens
2.3.1.26 C92A/C333A/C345A/C365A/C387A/C467A/C516A/C528A/C54 expressed in H5 cells or in ACAT-deficient Chinese hamster ovary cells: both mutants retain partial enzyme activity Homo sapiens
2.3.1.26 C92A/C333A/C345A/C365A/C387A/C467A/C516A/C528A/C54 this mutant is localized in endoplasmic reticulum in Chinese hamster ovary cells, homotetramer. Insensitive to p-chloromercuribenzene sulfonic acid Homo sapiens
2.3.1.26 C92A/C333A/C345A/C365A/C387A/C467A/C516A/C546A this mutant is localized in endoplasmic reticulum in Chinese hamster ovary cells Homo sapiens
2.3.1.26 C92A/C333A/C345A/C365A/C387A/C467A/C516A/C546A expressed in H5 cells or in ACAT-deficient Chinese hamster ovary cells: both mutants retain partial enzyme activity. Resistant to p-chloromercuribenzene sulfonic acid inhibition Homo sapiens
2.3.1.26 C92A/C333A/C345A/C365A/C387A/C516A/C528A/C546A sensitive to p-chloromercuribenzene sulfonic acid inhibition Homo sapiens
2.3.1.26 C92A/C333A/C345A/C365A/C467A/C516A/C546A expressed in H5 cells or in ACAT-deficient Chinese hamster ovary cells: both mutants retain partial enzyme activity. Resistant to p-chloromercuribenzene sulfonic acid inhibition Homo sapiens
2.3.1.26 C92A/C333A/C365A/C387A/C467A/C516A/C528A/C546A insensitive to p-chloromercuribenzene sulfonic acid Homo sapiens
2.3.1.26 C92A/C467A/C516A/C546A expressed in H5 cells or in ACAT-deficient Chinese hamster ovary cells: both mutants retain partial enzyme activity Homo sapiens
2.3.1.26 C92A/C467A/C516A/C546A this mutant is localized in endoplasmic reticulum in Chinese hamster ovary cells Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.1.26 5,5'-dithiobis-(2-nitrobenzoate) 50% inhibition of ACAT1 at 0.01 mM Homo sapiens
2.3.1.26 N-ethylmaleimide 50% inhibition of ACAT1 at 0.5 mM Homo sapiens
2.3.1.26 p-chloromercuribenzene sulfonic acid 50% inhibition of ACAT1 at 0.01 mM, Cys467 is one of the major targets for inhibition Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.26 additional information
-
 additional information for the various Cys-deficient mutant ACAT1s, the Km value for oleoyl-coenzyme A is approximately 2fold higher than the value for the wild-type enzyme, ranging between 0.012 und 0.015 mM Homo sapiens
2.3.1.26 0.0071
-
 oleoyl-CoA wild-type enzyme Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.1.26 50000
-
 immunoblotting, ACAT1 enzyme and Cys-mutants Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.26 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.26 of the recombinant enzyme using solubilization with CHAPS und KCl, chromatography on cobalt column and chromatography on monoclonal antibody affinity column Homo sapiens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.1.26 3.5 4 purified recombinant enzyme Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.26 oleoyl-CoA + cholesterol
-
 Homo sapiens CoA + cholesteryl oleate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.1.26 homotetramer use of chemical cross-linking agent disuccinimidyl suberate causes the formation of material two to four times the size of the monomeric enzyme when added to intact cells expressing either wild-type ACAT1 or mutant ACAT1 C92/333/345/365/387/467/516/528/546A Homo sapiens