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Literature summary for 2.3.1.26 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
Search for more literature for 2.3.1.26

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
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
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
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
C345A/C467A insensitive to p-chloromercuribenzene sulfonic acid Homo sapiens
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
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
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
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
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
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
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
C92A/C333A/C345A/C365A/C387A/C467A/C516A resistant to p-chloromercuribenzene sulfonic acid inhibition Homo sapiens
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
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
C92A/C333A/C345A/C365A/C387A/C467A/C516A/C546A this mutant is localized in endoplasmic reticulum in Chinese hamster ovary cells Homo sapiens
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
C92A/C333A/C345A/C365A/C387A/C516A/C528A/C546A sensitive to p-chloromercuribenzene sulfonic acid inhibition Homo sapiens
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
C92A/C333A/C365A/C387A/C467A/C516A/C528A/C546A insensitive to p-chloromercuribenzene sulfonic acid Homo sapiens
C92A/C467A/C516A/C546A expressed in H5 cells or in ACAT-deficient Chinese hamster ovary cells: both mutants retain partial enzyme activity Homo sapiens
C92A/C467A/C516A/C546A this mutant is localized in endoplasmic reticulum in Chinese hamster ovary cells Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
5,5'-dithiobis-(2-nitrobenzoate) 50% inhibition of ACAT1 at 0.01 mM Homo sapiens
N-ethylmaleimide 50% inhibition of ACAT1 at 0.5 mM Homo sapiens
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
0.0071
-
 oleoyl-CoA wild-type enzyme Homo sapiens

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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]

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

Substrates and Products (Substrate)

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

Subunits

Subunits Comment Organism
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