Information on EC 2.3.1.12 - dihydrolipoyllysine-residue acetyltransferase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
2.3.1.12
-
RECOMMENDED NAME
GeneOntology No.
dihydrolipoyllysine-residue acetyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
acetyl-CoA + enzyme N6-(dihydrolipoyl)lysine = CoA + enzyme N6-(S-acetyldihydrolipoyl)lysine
show the reaction diagram
reaction mechanism
-
acetyl-CoA + enzyme N6-(dihydrolipoyl)lysine = CoA + enzyme N6-(S-acetyldihydrolipoyl)lysine
show the reaction diagram
reaction mechanism
-
acetyl-CoA + enzyme N6-(dihydrolipoyl)lysine = CoA + enzyme N6-(S-acetyldihydrolipoyl)lysine
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Acyl group transfer
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
Citrate cycle (TCA cycle)
-
Glycolysis / Gluconeogenesis
-
Metabolic pathways
-
Microbial metabolism in diverse environments
-
pyruvate decarboxylation to acetyl CoA
-
Pyruvate metabolism
-
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:enzyme N6-(dihydrolipoyl)lysine S-acetyltransferase
A multimer (24-mer or 60-mer, depending on the source) of this enzyme forms the core of the pyruvate dehydrogenase multienzyme complex, and binds tightly both EC 1.2.4.1, pyruvate dehydrogenase (acetyl-transferring) and EC 1.8.1.4, dihydrolipoyl dehydrogenase. The lipoyl group of this enzyme is reductively acetylated by EC 1.2.4.1, and the only observed direction catalysed by EC 2.3.1.12 is that where the acetyl group is passed to coenzyme A.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
acetyltransferase, lipoate
-
-
-
-
DHLTA
-
-
-
-
dihydrolipoamide acetyltransferase
-
-
dihydrolipoamide acetyltransferase
-
-
dihydrolipoamide acetyltransferase
A6ZS09
E2 component of the mitochondrial pyruvate dehydrogenase complex
dihydrolipoate acetyltransferase
-
-
-
-
dihydrolipoic transacetylase
-
-
-
-
dihydrolipoyl acetyl transferase
-
-
dihydrolipoyl acetyltransferase
-
-
-
-
dihydrolipoyl acetyltransferase
-
-
dihydrolipoyl acetyltransferase
-
-
dihydrolipoyl acetyltransferase
-
glutathione-S-transferase fused to the inner lipoyl domain is used
dihydrolipoyl acetyltransferase
Q93T73
-
dihydrolipoyl acetyltransferase
-
-
dihydrolipoyl acetyltransferase component E2
-
-
dihydrolipoyl acetyltransferase E2p
-
-
dihydrolipoyl transacetylase
-
-
dihydrolipoyl transacetylase
-
is part of the human pyruvate dehydrogenase complex
dihydrolipoyl transacetylase
-
-
E2
-
-
-
-
E2p
-
-
-
-
lipoate acetyltransferase
-
-
-
-
lipoate transacetylase
-
-
-
-
lipoic acetyltransferase
-
-
-
-
lipoic acid acetyltransferase
-
-
-
-
lipoic transacetylase
-
-
-
-
myelin-proteolipid O-palmitoyltransferase
-
-
-
-
palmitoyl-CoA:myelin-proteolipid O-palmitoyltransferase
-
-
-
-
pyruvate dehydrogenase complex dihydrolipoamide acetyltransferase component
A6ZS09
-
thioltransacetylase A
-
-
-
-
transacetylase X
-
-
-
-
lipoylacetyltransferase
-
-
-
-
additional information
-
the enzyme is a subunit of the pyruvate dehydrogenase multienzyme complex
additional information
-
the enzyme forms the core unit E2 of the pyruvate dehydrogenase multienzyme complex binding the other components, i.e. pyruvate decarboxylase and dihydrolipoyl dehydrogenase, tightly at its peripheral domain
additional information
-
the enzyme forms the core unit E2 of the pyruvate dehydrogenase multienzyme complex binding the other components, i.e. pyruvate decarboxylase E1 and dihydrolipoyl dehydrogenase E3, tightly at its innerlipoyl or N-terminal lipoyl domain, respectively, composition overview
additional information
-
the enzyme is a subunit of the pyruvate dehydrogenase multienzyme complex, the enzyme forms the core unit E2 of the pyruvate dehydrogenase multienzyme complex binding the other components, i.e. pyruvate decarboxylase E1 and dihydrolipoyl dehydrogenase E3, tightly at its innerlipoyl or N-terminal lipoyl domain, respectively, composition overview
additional information
Q93T73
the enzyme is a subunit of the pyruvate dehydrogenase multienzyme complex
additional information
-
the enzyme is a subunit of the pyruvate dehydrogenase multienzyme complex, the enzyme forms the core unit E2 of the pyruvate dehydrogenase multienzyme complex binding the other components, i.e. pyruvate decarboxylase E1 and dihydrolipoyl dehydrogenase E3, tightly at its innerlipoyl or N-terminal lipoyl domain, respectively, composition overview
CAS REGISTRY NUMBER
COMMENTARY
9032-29-5
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
wild-type and mutants
-
-
Manually annotated by BRENDA team
broccoli
-
-
Manually annotated by BRENDA team
pigeon
-
-
Manually annotated by BRENDA team
K-12, wild-type and mutant strains
-
-
Manually annotated by BRENDA team
various wild-type and mutant strains
-
-
Manually annotated by BRENDA team
wild-type and mutants
-
-
Manually annotated by BRENDA team
Hansenula miso
-
-
-
Manually annotated by BRENDA team
enzyme forms the core of the pyruvate dehydrogenase multienzyme complex
-
-
Manually annotated by BRENDA team
cell lines BML1-ME, NIH3T3, PA317 and PE 501
-
-
Manually annotated by BRENDA team
lipoyl domain of strain 232; strains J, Beaufor, 232, Sue, YZ, and C1735/2, single copy gene pdhC, organized in the pdhCD operon
SwissProt
Manually annotated by BRENDA team
enzyme forms the core of the pyruvate dehydrogenase multienzyme complex
-
-
Manually annotated by BRENDA team
maize
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
Nov3r a lipoyl group-binding site inhibitor (related trifluoro-2-hydroxy-2-menthylpropionate compound) prevents pyruvate dehydrogenase kinase2 and GST-L2 (glutathione-S-transferase fused to the inner lipoyl domain (L2) of dihydrolipoyl acetyltransferase) binding and dissect the effects of Nov3r binding at the lipoyl group binding site on PDHK2 binding of other ligands
physiological function
-
in human pyruvate dehydrogenase complex the pyruvate dehydrogenase (E1) is bound to the E1-binding domain of dihydrolipoamide acetyltransferase (E2). K276 of hE2 is involved in the interaction with pyruvate dehydrogenase
physiological function
-
structural comparison by cryo-electronmicroscopy of the human full-length and truncated dihydrolipoyl acetyltransferase cores reveal flexible linkers emanating from the edges of trimers of the internal catalytic domains. Using the secondary structure constraints revealed the 8 A cryo-electronmicroscopy and the prokaryotic truncated dihydrolipoyl acetyltransferase atomic structure as a template, a pseudo atomic model of human truncated dihydrolipoyl acetyltransferase is derived. The active sites are conserved between the truncated prokaryotic and human enzyme. Marked structural differences are apparent in the hairpin domain and in the N-terminal helix connected to the flexible linker
physiological function
-
uing an in vitro reconstituted pyruvate dehydrogenase complex densitometry, isothermal titration calorimetry, and analytical ultracentrifugation evidence are provided that there are 40 copies of dihydrolipoyl transacetylase (E2p) and 20 copies of dihydrolipoamide dehydrogenase-binding protein (E3BP), in the E2p/E3BP core. The overall maximal stoichiometry of this in vitro assembled pyruvate dehydrogenase complex for dihydrolipoyl transacetylase: dihydrolipoamide dehydrogenase-binding protein: dihydrolipoamide dehydrogenase is 40:20:40:20
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + enzyme N6-(dihydrolipoyl)lysine
CoA + enzyme N6-(S-acetyldihydrolipoyl)lysine
show the reaction diagram
-
reductive acetylation of the enzyme
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
r
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
Q9SWR9
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
Hansenula miso
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
acetyl group is transferred from the S-acetyldihydrolipoyl moiety bound covalently to the transacetylase to CoA
-
r
additional information
?
-
-
protein bound acetyldihydrolipoyl moiety + CoA
-
-
-
additional information
?
-
-
E2 plays a central role in organization, integrated chemical reactions, and regulation of the multienzyme complex, overview, binding and activation of regulatory enzyme pyruvate dehydrogenase kinase isozymes PDK1-4, the enzyme mediates Ca2+-activation of isozymes of pyruvate dehydrogenase phosphatase severalfold, and enhances the accessibility of the E1 substrate for the regulatory enzymes, and mediate feedback effector control by NADH and acetyl-CoA, and modifies the allosteric control, mechanism, overview
-
-
-
additional information
?
-
-
the enzyme E2 has an enormous impact on pyruvate dehydrogenase kinase PDK phosphorylation of the pyruvate dehydrogenase E1 component by acting as a mobile binding framework and in facilitating and mediating regulation of PDK activity, isozyme PDK2 interacts very weakly with L2 domain of E2, but much tighter with the recombinant dimeric glutathione S-transferase-L2 domain fusion protein, importance of bifunctional binding, interaction of PDK2 with the lipoyl domains L1 and L2 of E2, and the E3-binding protein L3, overview
-
-
-
additional information
?
-
-
the enzyme mediates Ca2+-activation of isozymes 1 of pyruvate dehydrogenase phosphatase by 10fold, and enhances the accessibility of the E1 substrate for the regulatory enzymes, and mediate feedback effector control by NADH and acetyl-CoA, and modifies the allosteric control, mechanism, overview
-
-
-
additional information
?
-
-
interactions are different with the different isoforms of the regulatory enzymes of the multienzyme complex, signal mechanism for stimulation, overview, enzyme forms the core of the pyruvate dehydrogenase multienzyme complex, the flexibly held outer domains of the enzyme show dynamic, E2 is responsible for effector-modified interactions with the complex' regulatory enzymes pyruvate dehydrogenase kinase PDK and pyruvate dehydrogenase phosphatase PDP, which exist in different isoforms, overview
-
-
-
additional information
?
-
-
the overall complex reaction is irreversible
-
-
-
additional information
?
-
A6ZS09
Lat1 is a limiting longevity factor in nondividing cells. Overexpressing Lat1 extends cell survival during prolonged culture at stationary phase. Studies suggest that Lat1 overexpression extends life span by increasing metabolic fitness of the cell
-
-
-
additional information
?
-
-
in human pyruvate dehydrogenase complex the pyruvate dehydrogenase (E1) is bound to the E1-binding domain of dihydrolipoamide acetyltransferase (E2)
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
Q9SWR9
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
Hansenula miso
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
-
-
?
dihydrolipoamide + acetyl-CoA
S-acetyldihydrolipoamide + CoA
show the reaction diagram
-
acetyl group is transferred from the S-acetyldihydrolipoyl moiety bound covalently to the transacetylase to CoA
-
r
additional information
?
-
-
protein bound acetyldihydrolipoyl moiety + CoA
-
-
-
additional information
?
-
-
E2 plays a central role in organization, integrated chemical reactions, and regulation of the multienzyme complex, overview, binding and activation of regulatory enzyme pyruvate dehydrogenase kinase isozymes PDK1-4, the enzyme mediates Ca2+-activation of isozymes of pyruvate dehydrogenase phosphatase severalfold, and enhances the accessibility of the E1 substrate for the regulatory enzymes, and mediate feedback effector control by NADH and acetyl-CoA, and modifies the allosteric control, mechanism, overview
-
-
-
additional information
?
-
-
the enzyme E2 has an enormous impact on pyruvate dehydrogenase kinase PDK phosphorylation of the pyruvate dehydrogenase E1 component by acting as a mobile binding framework and in facilitating and mediating regulation of PDK activity, isozyme PDK2 interacts very weakly with L2 domain of E2, but much tighter with the recombinant dimeric glutathione S-transferase-L2 domain fusion protein, importance of bifunctional binding, interaction of PDK2 with the lipoyl domains L1 and L2 of E2, and the E3-binding protein L3, overview
-
-
-
additional information
?
-
-
the enzyme mediates Ca2+-activation of isozymes 1 of pyruvate dehydrogenase phosphatase by 10fold, and enhances the accessibility of the E1 substrate for the regulatory enzymes, and mediate feedback effector control by NADH and acetyl-CoA, and modifies the allosteric control, mechanism, overview
-
-
-
additional information
?
-
A6ZS09
Lat1 is a limiting longevity factor in nondividing cells. Overexpressing Lat1 extends cell survival during prolonged culture at stationary phase. Studies suggest that Lat1 overexpression extends life span by increasing metabolic fitness of the cell
-
-
-
additional information
?
-
-
in human pyruvate dehydrogenase complex the pyruvate dehydrogenase (E1) is bound to the E1-binding domain of dihydrolipoamide acetyltransferase (E2)
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
K+
-
association of the pyruvate dehydrogenase kinase2 and GST-L2 (glutathione-S-transferase fused to the inner lipoyl domain (L2) of dihydrolipoyl acetyltransferase (E2)) dimers is enhanced by K+
phosphate
-
phosphate has a pronounced effect in increasing ligand interference with pyruvate dehydrogenase kinase2 and GST-L2 (glutathione-S-transferase fused to the inner lipoyl domain (L2) of dihydrolipoyl acetyltransferase (E2))
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,2-diselenolane-3-valeric acid
-
competitive with respect to dihydrolipoamide and non-competitive with respect to acetyl-CoA
CoA
-
competitive to acetyl-CoA, non-competitive to dihydrolipoamide
guanidine hydrochloride
-
50% inhibition at 0.3 M, complete inhibition at 0.7-1 M
Iodine
-
complete inactivation at 2.5 mol iodine per mol of subunit
N-Acetylimidazole
-
modifies tyrosyl residues and dissociates the enzyme into subunits
palmitoyl-CoA
-
competitive to acetyl-CoA, non-competitive to dihydrolipoamide
S-acetyldihydrolipoamide
-
competitive to dihydrolipoamide, non-competitive to acetyl-CoA
lipoamidase
-
complete loss of activity after 3 h
-
additional information
-
starvation inhibits the overall complex reaction
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.013
-
acetyl-CoA
-
catalytic domain after expression in Escherichia coli
0.022
-
acetyl-CoA
-
wild-type enzyme
0.028
-
acetyl-CoA
-
S558A mutant
0.03
0.1
acetyl-CoA
-
-
0.036
-
acetyl-CoA
-
H610C mutant
0.052
-
acetyl-CoA
-
N614D mutant
0.13
0.4
Dihydrolipoamide
-
-
1.08
-
Dihydrolipoamide
-
N614D mutant
1.2
-
Dihydrolipoamide
-
catalytic domain after expression in Escherichia coli
2.2
-
Dihydrolipoamide
-
-
4.2
-
Dihydrolipoamide
-
-
5.3
-
Dihydrolipoamide
-
H610C mutant
5.37
-
Dihydrolipoamide
-
wild-type enzyme
6.4
-
Dihydrolipoamide
-
S558A mutant
additional information
-
additional information
-
-
-
additional information
-
additional information
-
the complex assembly and maintainance is regulated by thermodynamic homeostasis or entropy-enthalpy compensation
-
additional information
-
additional information
-
complex component interactions among each other and with regulatory enzyme pyruvate dehydrogenase kinase isozyme 2, overview, dissociation constants of interaction of L2 and recombinant GST-L2 with PDK isozyme 2, reduced PDK2 binding to E2-E1, reversible bifunctional binding to L2 with the mandatory singly held transition fits the proposed 'hand-over-hand' movement of a kinase dimer to access E1 without dissociating from the complex
-
additional information
-
additional information
-
binding constants of multienzyme complex components and regulatory enzymes, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00417
-
acetyl-CoA
-
with dihydrolipoamide, H610C mutant
0.0143
-
acetyl-CoA
-
with dihydrolipoamide, S558A mutant
0.282
-
acetyl-CoA
-
with dihydrolipoamide, N614D mutant
2.4
-
acetyl-CoA
-
with dihydrolipoamide, wild-type enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.89
-
-
fusion protein with glutathione S-transferase
4.09
-
-
after removal of glutathione S-transferase moiety
19.4
-
-
-
19.7
-
-
-
34.1
-
-
purified 1-lip E2
47.1
-
-
catalytic domain
119
-
-
catalytic domain
191
-
Q9SWR9
-
additional information
-
-
specific activities in various wild-type and mutant strains
additional information
-
-
specific activities of fragments after protease cleavage
additional information
-
-
specific activity of native enzyme and proteolytic fragments in various experiments
additional information
-
-
-
additional information
-
-
specific activity after growth on various carbon sources
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.8
8.5
-
-
8
-
-
assay at
additional information
-
-
reactivity depending on buffer system
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
-
25
-
-
assay at
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
pyruvate dehydrogenase complex bound to inner mitochondrial membrane
Manually annotated by BRENDA team
-
pyruvate dehydrogenase complex bound to inner mitochondrial membrane
Manually annotated by BRENDA team
-
wild-type enzyme is soluble, whereas mutant enzyme with deletion of all three lipoyl domains aggregates
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Pyrobaculum aerophilum (strain ATCC 51768 / IM2 / DSM 7523 / JCM 9630 / NBRC 100827)
Pyrobaculum aerophilum (strain ATCC 51768 / IM2 / DSM 7523 / JCM 9630 / NBRC 100827)
Pyrobaculum aerophilum (strain ATCC 51768 / IM2 / DSM 7523 / JCM 9630 / NBRC 100827)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30000
-
-
gel filtration in the presence of guanidinium hydrochloride
46930
-
Q9SWR9
calculation from nucleotide sequence
50080
-
-
calculation from nucleotide sequence
51000
-
-
gel filtration
52000
53000
Q9SWR9
SDS-PAGE
58000
59000
-
SDS-PAGE
58890
-
-
calculation from nucleotide sequence
59000
-
-
SDS-PAGE
60000
-
-
calculation from nucleotide sequence
60000
-
-
mature protein, SDS-PAGE
60100
64500
-
sedimentation equilibrium, depending on buffer
63000
-
-
sedimentation equilibrium
64260
-
-
calculation from nucleotide sequence
64910
-
-
calculation from nucleotide sequence
65960
-
-
calculation from nucleotide sequence
67000
-
-
plastid precursor protein, SDS-PAGE
68000
-
-
SDS-PAGE
70000
-
-
SDS-PAGE
70000
-
-
SDS-PAGE
73000
75000
-
SDS-PAGE
74000
-
-
SDS-PAGE
74000
-
-
SDS-PAGE and sedimentation equilibrium data
78000
-
-
SDS-PAGE
80000
-
-
SDS-PAGE
80000
-
-
SDS-PAGE and calculated from amino acid sequence
80000
-
-
SDS-PAGE
81000
-
-
SDS-PAGE, gel filtration
83000
-
-
SDS-PAGE
83000
-
-
wild-type enzyme, SDS-PAGE
127000
-
-
fusion protein of 11000 Da fragment with beta-galactosidase, SDS-PAGE
150000
-
-
no component of pyruvate dehydrogenase complex, sucrose density gradient centrifugation
200000
-
-
sedimentation equilibrium centrifugation, light scattering, sedimentation and diffusion coefficients
200000
-
-
at 1.8 - 2.8 M guanidine hydrochloride, complex dissociates at higher guanidine hydrochloride levels to a monomeric form with MW 82000
500000
-
-
above, gel filtration; gel filtration
530000
-
-
gel filtration
670000
-
-
gel filtration
864000
-
-
crystallization experiments
1000000
-
-
sedimentation equilibrium centrifugation
1470000
-
-
sedimentation equilibrium experiments in 0.83 M acetic and and 0.005 M sodium chloride
1548000
-
-
native enzyme
1560000
-
-
fluorescence anisotropy decay experiments
1800000
1970000
-
calculation from sedimentation and diffusion constants, meniscus depletion method
2700000
-
Q9SWR9
gel filtration
2800000
-
-
calculation from subunit composition of pyruvate dehydrogenase complex
3100000
-
-
-
additional information
-
-
molecular weight of lipolyl domains
additional information
-
-
molecular weight of lipolyl domains
additional information
-
-
molecular weight of proteolytic fragments
additional information
-
-
molecular weight of fragments after treatment with trypsin
additional information
-
-
molecular weight of proteolytic fragments
additional information
-
-
molecular weight of proteolytic fragments
additional information
-
-
molecular weight of lipolyl domains
additional information
-
-
molecular weight of proteolytic fragments
additional information
-
-
molecular weight of proteolytic fragments
additional information
-
-
molecular weight of E2 complexed with E1 or E3
additional information
-
-
molecular weight of proteolytic fragments
additional information
-
-
high molecular mass species in the absence of guanidinium hydrochloride
additional information
-
-
molecular weight of proteolytic fragments
additional information
-
-
molecular weight of proteolytic fragments
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
Q93T73
x * 33100, about, E2, sequence calculation
?
-
x * 45953, recombinant 1-lip E2, mass spectrometry, x * 8982, recombinant unacetylated hybrid lipoyl domain, mass spectrometry, x * 9019, recombinant fully acetylated hybrid lipoyl domain, mass spectrometry
?
-
x * 59000, SDS-PAGE, recombinant protein
dimer
-
multimer formation is probably lost by loss of a small segment during genetic rearrangement
dimer
-
2 * 36000, in the presence of dilute acetic acid
dimer
-
60mer, analytical ultracentrifugation
polymer
-
stoichiometry of pyruvate dehydrogenase complexes
polymer
-
24 * 48000
polymer
-
60 * 52000
polymer
-
stoichiometry of pyruvate dehydrogenase complexes
polymer
-
22 or 24 subunits
polymer
-
24 * 65959, calculation from nucleotide sequence
polymer
-
x * 46265, calculation from nucleotide sequence
polymer
-
domain structure
polymer
-
24 * 60000, SDS-PAGE; x * 60000, SDS-PAGE
polymer
-
x * 48546, calculation from nucleotide sequence
polymer
-
24 * 27500, SDS-PAGE, light scattering experiments, quarternary structure
polymer
-
-
polymer
-
primary structure of lipoyl domain
polymer
-
30 * 66000, SDS-PAGE, sedimentation equilibrium
polymer
-
domain structure of subunits: lipoyl domain MW 28000, subunit binding domain MW 26000, sedimentation equilibrium
polymer
-
30 * 70000, SDS-PAGE, each polypeptide chains has 2 domains
polymer
-
gel filtration in presence of guanidine-HCl, anomalous migration on SDS
polymer
-
-
polymer
-
24 identical subunits, data from crystallographic, biochemical and electron microscopic methods
polymer
-
24 * 74000 SDS-PAGE, gel filtration in 6 M guanidine-HCl, sedimentation equilibrium in 6 M guanidine-HCl
polymer
-
24 * 40000, linked by noncovalent bonds
polymer
-
24 * 36000, linked by noncovalent bonds
polymer
-
60 polypeptide chains, crystal structure
polymer
-
stoichiometry of pyruvate dehydrogenase complexes
polymer
-
60 subunits
polymer
-
60 * 42000, SDS-PAGE
polymer
-
60 subunits; stoichiometry of pyruvate dehydrogenase complexes
polymer
-
60 * 52000; stoichiometry of pyruvate dehydrogenase complexes
trimer
-
3 * 82000, gel filtration, trimeric form occurs in solutions with 4 M guanidine hydrochloride
dimer
-
GST-L2, glutathione-S-transferase fused to the inner lipoyl domain (L2) of dihydrolipoyl acetyltransferase exists as a dimer
additional information
-
treatment with NaCl results in 2 fractions with E2 activity, MW 55000 and MW 78000, SDS-PAGE, gel filtration
additional information
-
enzyme forms stable aggregates
additional information
-
the enzyme forms the core unit E2, consisting of 4 domains in 60mer, a trimer of 3 20mers, of the pyruvate dehydrogenase multienzyme complex binding the other components, i.e. pyruvate decarboxylase and dihydrolipoyl dehydrogenase, tightly at its innerlipoyl or N-terminal lipoyl domain, respectively, composition overview
additional information
-
the enzyme forms the core unit of the pyruvate dehydrogenase multienzyme complex binding the other components, i.e. pyruvate decarboxylase E1 and dihydrolipoyl dehydrogenase E3, tightly at its peripheral domain, Arg135 is important for interactions with E1 and E3, Met131 is involved in binding of E1, interaction analysis by surface plasmon resonance
additional information
-
E2 is composed of 4 domains including the innerlipoyl domain L2 important for binding of E1
additional information
-
enzyme is a 60mer, E2 domain structure: 4 domains, i.e. L1, L2, B, and I domain, connected by linker oligomers, overview, overall multienzyme complex organization, overview
additional information
-
evidence for a novel subunit organization in which dihydrolipoamide dehydrogenase E3 and E3BP form subcomplexes with a 1:2 stoichiometry implying the existence of a network of dihydrolipoamide dehydrogenase cross-bridges linking pairs of E3-binding proteins across the surface of the dihydrolipoamide acetyltransferase E2 core assembly. One of the E3-binding protein lipoyl domains docks into the dihydrolipoamide dehydrogenase E3 active site
additional information
-
cryoelectron microscopic analysis of the reconstructed three-dimensional structure of the purified E2E3 complex (dihydrolipoyl acetyltransferase/dihydrolipoyl dehydrogenase) and use of automated docking methods to interpret the density map in terms of the probable localization of the dihydrolipoyl acetyltransferase E2 and dihydrolipoyl dehydrogenase E3 molecules. The arrangement of pyruvate decarboxylase E1 and dihydrolipoyl dehydrogenase E3 molecules in the outer shell of the pyruvate dehydrogenase complex are remarkably similar and indicate that the design of the annular gap allows the lipoyl domain to have access to the active sites of pyruvate decarboxylase E1, dihydrolipoyl acetyltransferase E2, and dihydrolipoyl dehydrogenase E3 enzymes from within the annular gap
additional information
-
distinct modes of recognition of the lipoyl domain of the dihydrolipoyl acetyltransferase (E2) component as substrate by the E1 and E3 components of the pyruvate dehydrogenase multienzyme complex
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
lipoprotein
-
dynamic properties of lipoyl domains
lipoprotein
-
30 polypeptide chains with 2 lipoyl domains each
lipoprotein
-
-
lipoprotein
-
3 lipoyl domains
lipoprotein
-
3 lipoyl domains separated by regions rich in alanine and proline
lipoprotein
-
-
lipoprotein
-
1 molecule lipoic acid per polypeptide chain
lipoprotein
-
2 lipoyl domains
lipoprotein
-
-
lipoprotein
-
1 lipoyl domain
lipoprotein
-
2 lipoyl domains per polypeptide chain
lipoprotein
-
1 lipoyl domain
lipoprotein
-
-
lipoprotein
-
23 mol of lipoic acid per mol of enzyme
lipoprotein
Q9SWR9
1 lipoyl domain
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hanging drop vapor diffusion method
-
pyruvate dehydrogenase complex
-
E2-E3 subcomplex of pyruvate dehydrogenase, E2: EC 2.3.1.12, E3: EC 1.8.1.4, various crystallization conditions
-
E3 dimer with E2 and a small binding domain E2BD, hanging drop vapor diffusion method
-
crystal structure of pyruvate dehydrogenase kinase 2 bound to the inner lipoyl-bearing domain of dihydrolipoamide transacetylase is determined. Crystal structure reveals a pyruvate dehydrogenase kinase 2 dimer complexed with two inner lipoyl-bearing domains of dihydrolipoamide transacetylase. Comparison with apo-pyruvate dehydrogenase kinase 2 shows that dihydrolipoamide transacetylase binding causes rearrangements in PDHK2 structure that affect the dihydrolipoamidetransacetylase- and pyruvate dehydrogenase-binding sites
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3
11
-
isolated E. coli lipoyl domains stay soluble in this range
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
60
-
-
stable up to
70
-
-
1 h stable, isolated lipoyl domains
70
-
-
incubation converts core of the enzyme into an unidentified active molecular form X, loss of 10% enzyme activity after 5 h
78
-
-
temperatures below result in less than 5% loss in enzyme activity
80
-
-
partly precipitation above
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
sensitive to proteolysis
-
sensitive to proteolysis
-
freezing and thawing does not cause loss of activity
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
liquid nitrogen, 50 mM potassium phosphate buffer pH 7.0, 0.5 mM EDTA, 0.05 mM phenylmethylsulfonylfluoride
-
-18°C, 0.05 mM potassium phosphate buffer, pH 7.0, 0.5 mM EDTA, more than 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
fusion protein with glutathione S-transferase
-
catalytic domain mutants
-
fusion protein of 6 amino acids form beta-galactosidase, the papa-4 region and the catalytic domain of E2
-
proteolytic fragments, isolation of lipoyl domain and catalytic domain
-
pyruvate dehydrogenase complex, composed of EC 1.2.4.1, EC 1.8.1.4, EC 2.3.1.12
-
from pyruvate dehydrogenase complex
-
preparation of E2-X subcomplex: E2 i.e. EC 2.3.1.12, X i.e. component X of mammalian pyruvate dehydrogenase complex
-
purification of tryptic fragments
-
pyruvate dehydrogenase complex, composed of EC 1.2.4.1, EC 1.8.1.4, EC 2.3.1.12
-
purification from pyruvate dehydrogenase complex, composed of EC 1.2.4.1, EC 1.8.1.4, EC 2.3.1.12
-
E1-E2 complex
-
fusion protein of 6 amino acids form beta-galactosidase, the apa-4 region and the catalytic domain of E2
-
isolation of lipoyl domains
-
pyruvate dehydrogenase complex and tryptic fragments of E2
-
pyruvate dehydrogenase complex, stoichiometry E1:E2:E3 is 1.56:1:0.89
-
recombinant 1-lip E2 and recombinant hybrid lipoyl domain by ammonium sulfate fractionation, ion exchange and hydrophobic interaction chromatography
-
wild-type and mutants with deletions of lipolyl domains
-
peptides after limited proteolysis
-
purification of pyruvate dehydrogenase complex, composed of EC 1.2.4.1, EC 1.8.1.4, EC 2.3.1.12
Hansenula miso
-
highly
-
purification of the multienzyme complex
-
using a sephacryl S-400 HR column
-
using Ni-nitrilotriacetate-agarose affinity chromatography
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
Q93T73
pyruvate dehydrogenase complex, composed of EC 1.2.4.1, EC 1.8.1.4, EC 2.3.1.12
-
from pyruvate dehydrogenase complex
-
purification of the multienzyme complex
-
pyruvate dehydrogenase complex, composed of EC 1.2.4.1, EC 1.8.1.4. EC 2.3.1.12
-
E2-E3 subcomplex of pyruvate dehydrogenase, i.e. dihydrolipoamide acetyltransferase and dehydrogenase
-
-
Q9SWR9
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
wild-type and deletion mutants, expression in Escherichia coli
-
wild-type and mutants
-
functional overexpression of the enzyme E2 with a single hybrid lipoyl domain per subunit, i.e. 1-lip E2, in strain JRG1342 and BL21(DE3), overexpression of the isolated hybrid lipoyl domain in strain JM 101
-
wild-type and mutants
-
catalytic domain, residues 173-427
-
expression of subgene for lipoyl domain in Escherichia coli
-
overexpression of truncated thrombin-cleavable enzyme in Escherichia coli strain BL21(DE3)
-
a recombinant fragment hL2S (containing the second lipoyl domain (L2), second hinge region, E1-binding domain (S) and third hinge region of hE2, residues 128–330) is overexpressed in Escherichia coli
-
expressed in Escherichia coli
-
expression of the enzymes' L2 domain as GST-fusion protein
-
inner lipoyl domain (L2) of dihydrolipoyl acetyltransferase is expressed as a GST fusion protein
-
the inner lipoyl-bearing domain of dihydrolipoamide transacetylase is expressed as a GST fusion protein in Escherichia coli
-
knock-out mutants
-
gene pdhC, organized in the pdhCD operon, physical map, DNA and amino acid sequence determination and analysis, expression as His6-tagged enzyme in Escherichia coli
Q93T73
-
Q9SWR9
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
K96A
-
mutation in potential lipoylation site, mutant protein entirely supports the assignment of Lys96 as the site of lipoylation
G610C
-
500fold reduced enzymatic activity
N614D
-
9fold reduced enzymatic activity
S558A
-
200fold reduced enzymatic activity
K244Q
-
unlipoylatable domain
K136A
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
K153A
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
M131A
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
R135A
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
R135C
-
site-directed mutagenesis, additional alkylation of the mutant enzymes with methyl, ethyl, propyl and butyl groups, the modifications cause alterations in interaction of core unit, E1 and E3 in the enzyme complex compared to the wild-type complex, thermodynamics and kinetics in comparison to the wild-type enzyme, overview
R135K
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
R135L
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
R135M
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
R139A
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
R146A
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
R156A
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
S133A
-
site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme
A174S
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 69%
D164A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 55%
D172A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 66%
E162A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 93%
E179A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 28%
E182A
-
binding of mutant L2 domain to pyruvate dehydrogenase phosphatase isozyme 1 is hindered
E182A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 100%
E182Q
-
binding of mutant L2 domain to pyruvate dehydrogenase phosphatase isozyme 1 is hindered
E183A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 98%
F231A
-
ability to be post-translationally lipoylated remains
I176A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: not measurable
I229A
-
ability to be post-translationally lipoylated remains
I229A/F231A
-
ability to be post-translationally lipoylated remains, far-UV CD spectrum differs from wild-type enzyme
K173A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: not measurable
K276A
-
mutant shows negligible binding to human pyruvate dehydrogenase with 86fold higher KD compared to wild-type
L140A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: not measurable
L189A
-
ability to be post-translationally lipoylated remains
M194A
-
ability to be post-translationally lipoylated remains
R196A
-
mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 79%
R297A
-
mutant is found to have KD 6.8fold higher than that for the wild-type, indicating a possible involvement of this residue in the interaction with human pyruvate dehydrogenase, but not with the C-terminal residue of beta-subunit
V180S/E181L
-
binding of mutant L2 domain to pyruvate dehydrogenase phosphatase isozyme 1 is hindered
K96R
-
mutation in potential lipoylation site, mutant protein entirely supports the assignment of Lys96 as the site of lipoylation
additional information
-
protein includes a single lipoyl-prosthetic group covalently attached to Lys96. Despite low primary sequence identity with the bacterial enzyme, the plant protein is recognized and modified by Escherichia coli E2 lipoyl-addition system
M197A
-
ability to be post-translationally lipoylated remains
additional information
-
construction of diverse L2 domain mutants, e.g. substitutions of A172, D173, Leu140, Glu162, Glu181, and Glu179 highly reduce binding of L2 domain to pyruvate dehydrogenase phosphatase isozyme 1
additional information
-
a truncated version of dihydrolipoyl acetyltransferase is created lacking the N-terminal 319 amino acids
V188A
-
ability to be post-translationally lipoylated remains, far-UV CD spectrum differs from wild-type enzyme
additional information
A6ZS09
deleting the LAT1 gene abolishes life span extension induced by calorie restriction. Overexpressing Lat1 extends life span, and this life span extension is not further increased by calorie restriction. Similar to calorie restriction, life span extension by Lat1 overexpression largely requires mitochondrial respiration. Lat1 overexpression does not require the Sir2 family to extend life span, suggesting that Lat1 mediates a branch of the calorie restriction pathway that functions in parallel to the Sir2 family
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
reconstitution with components from Escherichia coli and Azotobacter vinelandii, optimum catalytic stoichiometry of E1:E2:E3 is 1:1:0.5
-
reconstitution of pyruvate dehydrogenase complex
-
reconstitution of E1 with E2-E3 complex with a stoichiometry of E1:E2:E3 of 2:1:1
-
reconstitution of pyruvate dehydrogenase complex, maximum activity is produced when transacetylase accommodates 12 pyruvate dehydrogenase dimers and 6 flavoprotein dimers
-
reconstitution of pyruvate dehydrogenase complex, stoichiometry pyruvate dehydrogenase:transacetylase:flavoprotein is 1:0.35:0.4
-
reconstitution of the active multienzyme complex with recombinant components
-
treatment with dilute acetic acid solution results in dissociation into inactive subunits with MW 70000, removal of the acid results in restoration of enzymatic activity
-
treatment with guanidine hydrochloride and its subsequent removal results in little recovery of the core, but full recovery of X
-
treatment with guanidine hydrochloride results in dissociation into subunits, removal of guanidine hydrochloride results in refolding of the enzyme with 95% of the original activity
-
treatment with guanidine hydrochloride and its subsequent removal results in refolding
-
reconstitution of pyruvate dehydrogenase complex
-
reconstitution of pyruvate dehydrogenase complex with components from Escherichia coli and Salmonella typhimurium
-
reconstitution of pyruvate dehydrogenase complex, optimum catalytic stoichiometry of E1:E2:E3 is 5.2:11.5:2
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
molecular biology
A6ZS09
dihydrolipoamide acetyltransferase is shown to be a metabolic longevity factor and is required for calorie restriction-mediated life span extension