DB code: M00189

RLCP classification 3.1177.805.79 : Transfer
CATH domain 2.40.50.100 : OB fold (Dihydrolipoamide Acetyltransferase, E2P)
2.40.50.100 : OB fold (Dihydrolipoamide Acetyltransferase, E2P)
2.40.50.100 : OB fold (Dihydrolipoamide Acetyltransferase, E2P)
-.-.-.- :
4.10.320.10 : Dihydrolipoamide Transferase
3.30.559.10 : Chloramphenicol Acetyltransferase Catalytic domain
E.C. 2.3.1.12
CSA
M-CSA
MACiE

CATH domain Related DB codes (homologues)
2.40.50.100 : OB fold (Dihydrolipoamide Acetyltransferase, E2P) M00163 M00222 M00145 M00188 T00223 M00190 M00191 M00208
3.30.559.10 : Chloramphenicol Acetyltransferase M00188 T00223 M00190 M00191
4.10.320.10 : Dihydrolipoamide Transferase M00188 T00223 M00190 M00191

Uniprot Enzyme Name
UniprotKB Protein name Synonyms Pfam
P10802 Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex
EC 2.3.1.12
E2
Dihydrolipoamide acetyltransferase component of pyruvate dehydrogenase complex
PF00198 (2-oxoacid_dh)
PF00364 (Biotin_lipoyl)
PF02817 (E3_binding)
[Graphical View]

KEGG enzyme name
dihydrolipoyllysine-residue acetyltransferase
acetyl-CoA:dihydrolipoamide S-acetyltransferase
dihydrolipoamide S-acetyltransferase
dihydrolipoate acetyltransferase
dihydrolipoic transacetylase
dihydrolipoyl acetyltransferase
lipoate acetyltransferase
lipoate transacetylase
lipoic acetyltransferase
lipoic acid acetyltransferase
lipoic transacetylase
lipoylacetyltransferase
thioltransacetylase A
transacetylase X
enzyme-dihydrolipoyllysine:acetyl-CoA S-acetyltransferase
acetyl-CoA:enzyme 6-N-(dihydrolipoyl)lysine S-acetyltransferase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P10802 ODP2_AZOVI Acetyl-CoA + enzyme N(6)-(dihydrolipoyl)lysine = CoA + enzyme N(6)-(S-acetyldihydrolipoyl)lysine. Forms a 24-polypeptide structural core with octahedral symmetry. Binds 3 lipoyl cofactors covalently (By similarity).

KEGG Pathways
Map code Pathways E.C.
MAP00010 Glycolysis / Gluconeogenesis
MAP00020 Citrate cycle (TCA cycle)
MAP00620 Pyruvate metabolism

Compound table
Substrates Products Intermediates
KEGG-id C00010 L00017 C00024 C15973
E.C.
Compound CoA Enzyme N(6)-(S-acetyldihydrolipoyl)lysine Acetyl-CoA Enzyme N(6)-(dihydrolipoyl)lysine Tetrahedral intermediate
Type amine group,carbohydrate,nucleotide ,peptide/protein,sulfhydryl group amide group,carbohydrate,lipid,peptide/protein,sulfhydryl group,sulfide group amine group,carbohydrate,nucleotide ,peptide/protein,sulfide group amide group,lipid,peptide/protein,sulfhydryl group
ChEBI 15346
 15346
 		15351
 15351
PubChem 6816
 87642
 6816
 87642
 		444493
 6302
 444493
 6302
1iyuA Unbound Unbound Unbound Unbound Unbound
1iyvA Unbound Unbound Unbound Unbound Unbound
1dpbA Unbound Unbound Unbound Unbound Unbound
1dpcA Unbound Unbound Unbound Unbound Unbound
1dpdA Unbound Unbound Unbound Unbound Unbound
1eaaA Unbound Unbound Unbound Unbound Unbound
1eabA Bound:COA Unbound Unbound Analogue:LPM Unbound
1eacA Bound:CAO Unbound Unbound Analogue:DTT Unbound
1eadA Analogue:CAO Unbound Unbound Unbound Unbound
1eaeA Unbound Unbound Unbound Analogue:LPM Unbound
1eafA Unbound Unbound Unbound Unbound Intermediate-analogue:SO3

Reference for Active-site residues
resource references E.C.
Swiss-prot;P10802

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1iyuA LYS 39(Lipolyl binding)
1iyvA LYS 39(Lipolyl binding)
1dpbA SER 558;;ASN 614 mutant H610C
1dpcA SER 558;HIS 610; HIS 610 mutant N614D
1dpdA ;HIS 610;ASN 614 HIS 610 mutant S558A
1eaaA SER 558;HIS 610;ASN 614 HIS 610
1eabA SER 558;HIS 610;ASN 614 HIS 610
1eacA SER 558;HIS 610;ASN 614 HIS 610
1eadA SER 558;HIS 610;ASN 614 HIS 610
1eaeA SER 558;HIS 610;ASN 614 HIS 610
1eafA SER 558;HIS 610;ASN 614 HIS 610

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[13]
p.1547
[14]
Scheme I, p.3899-3901
[17]
p.1194-1195
[20]
Fig.1, p.4292-4295
[32]
Fig.6

References
[1]
Resource
Comments
Medline ID
PubMed ID 389239
Journal Biochem Biophys Res Commun
Year 1979
Volume 90
Pages 431-8
Authors Fuller CC, Reed LJ, Oliver RM, Hackert ML
Title Crystallization of a dihydrolipoyl transacetylase--dihydrolipoyl dehydrogenase subcomplex and its implications regarding the subunit structure of the pyruvate dehydrogenase complex from Escherichia coli.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 6376124
Journal Eur J Biochem
Year 1984
Volume 141
Pages 361-74
Authors Spencer ME, Darlison MG, Stephens PE, Duckenfield IK, Guest JR
Title Nucleotide sequence of the sucB gene encoding the dihydrolipoamide succinyltransferase of Escherichia coli K12 and homology with the corresponding acetyltransferase.
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID 3101735
Journal Biochemistry
Year 1986
Volume 25
Pages 8173-8
Authors Yang YS, Frey PA
Title Dihydrolipoyl transacetylase of Escherichia coli. Formation of 8-S-acetyldihydrolipoamide.
Related PDB
Related UniProtKB
[4]
Resource
Comments LIPOYL DOMAIN CONFORMATION.
Medline ID 89052887
PubMed ID 3191993
Journal FEBS Lett
Year 1988
Volume 240
Pages 205-10
Authors Hanemaaijer R, Vervoort J, Westphal AH, de Kok A, Veeger C
Title Mobile sequences in the pyruvate dehydrogenase complex, the E2 component, the catalytic domain and the 2-oxoglutarate dehydrogenase complex of Azotobacter vinelandii, as detected by 600 MHz 1H-NMR spectroscopy.
Related PDB
Related UniProtKB P10802
[5]
Resource
Comments
Medline ID
PubMed ID 2271545
Journal Biochemistry
Year 1990
Volume 29
Pages 8614-9
Authors Niu XD, Stoops JK, Reed LJ
Title Overexpression and mutagenesis of the catalytic domain of dihydrolipoamide acetyltransferase from Saccharomyces cerevisiae.
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID 2192914
Journal FEBS Lett
Year 1990
Volume 264
Pages 206-10
Authors Dardel F, Packman LC, Perham RN
Title Expression in Escherichia coli of a sub-gene encoding the lipoyl domain of the pyruvate dehydrogenase complex of Bacillus stearothermophilus.
Related PDB
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID 1908777
Journal Eur J Biochem
Year 1991
Volume 200
Pages 29-34
Authors Schulze E, Benen JA, Westphal AH, de Kok A
Title Interaction of lipoamide dehydrogenase with the dihydrolipoyl transacetylase component of the pyruvate dehydrogenase complex from Azotobacter vinelandii.
Related PDB
Related UniProtKB
[8]
Resource
Comments
Medline ID
PubMed ID 1935951
Journal Eur J Biochem
Year 1991
Volume 201
Pages 561-8
Authors Schulze E, Westphal AH, Obmolova G, Mattevi A, Hol WG, de Kok A
Title The catalytic domain of the dihydrolipoyl transacetylase component of the pyruvate dehydrogenase complex from Azotobacter vinelandii and Escherichia coli. Expression, purification, properties and preliminary X-ray analysis.
Related PDB
Related UniProtKB
[9]
Resource
Comments
Medline ID
PubMed ID 1590756
Journal Biochem J
Year 1992
Volume 283
Pages 665-71
Authors Hipps DS, Perham RN
Title Expression in Escherichia coli of a sub-gene encoding the lipoyl and peripheral subunit-binding domains of the dihydrolipoamide acetyltransferase component of the pyruvate dehydrogenase complex of Bacillus stearothermophilus.
Related PDB
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 1730230
Journal Eur J Biochem
Year 1992
Volume 203
Pages 245-50
Authors Snoep JL, Westphal AH, Benen JA, Teixeira de Mattos MJ, Neijssel OM, de Kok A
Title Isolation and characterisation of the pyruvate dehydrogenase complex of anaerobically grown Enterococcus faecalis NCTC 775.
Related PDB
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 1429691
Journal J Biol Chem
Year 1992
Volume 267
Pages 23484-8
Authors Green JD, Perham RN, Ullrich SJ, Appella E
Title Conformational studies of the interdomain linker peptides in the dihydrolipoyl acetyltransferase component of the pyruvate dehydrogenase multienzyme complex of Escherichia coli.
Related PDB
Related UniProtKB
[12]
Resource
Comments
Medline ID
PubMed ID 1589018
Journal Nature
Year 1992
Volume 357
Pages 196-7
Authors DeRosier DJ
Title Enzyme complexes. A farewell to arms.
Related PDB
Related UniProtKB
[13]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS) OF 381-637.
Medline ID 92196586
PubMed ID 1549782
Journal Science
Year 1992
Volume 255
Pages 1544-50
Authors Mattevi A, Obmolova G, Schulze E, Kalk KH, Westphal AH, de Kok A, Hol WG
Title Atomic structure of the cubic core of the pyruvate dehydrogenase multienzyme complex.
Related PDB 1eaa 1eab 1eac 1ead 1eae 1eaf
Related UniProtKB P10802
[14]
Resource
Comments
Medline ID
PubMed ID 8471601
Journal Biochemistry
Year 1993
Volume 32
Pages 3887-901
Authors Mattevi A, Obmolova G, Kalk KH, Teplyakov A, Hol WG
Title Crystallographic analysis of substrate binding and catalysis in dihydrolipoyl transacetylase (E2p).
Related PDB
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID 8436118
Journal Eur J Biochem
Year 1993
Volume 211
Pages 591-9
Authors Schulze E, Westphal AH, Hanemaaijer R, de Kok A
Title Structure/function relationships in the pyruvate dehydrogenase complex from Azotobacter vinelandii. Role of the linker region between the binding and catalytic domain of the dihydrolipoyl transacetylase component.
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID 8500617
Journal FEBS Lett
Year 1993
Volume 323
Pages 243-6
Authors Machado RS, Guest JR, Williamson MP
Title Mobility in pyruvate dehydrogenase complexes with multiple lipoyl domains.
Related PDB
Related UniProtKB
[17]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 8487300
Journal J Mol Biol
Year 1993
Volume 230
Pages 1183-99
Authors Mattevi A, Obmolova G, Kalk KH, Westphal AH, de Kok A, Hol WG
Title Refined crystal structure of the catalytic domain of dihydrolipoyl transacetylase (E2p) from Azotobacter vinelandii at 2.6 A resolution.
Related PDB 1dpb 1dpc 1dpd
Related UniProtKB
[18]
Resource
Comments
Medline ID
PubMed ID 8433963
Journal Protein Eng
Year 1993
Volume 6
Pages 101-8
Authors Turner SL, Russell GC, Williamson MP, Guest JR
Title Restructuring an interdomain linker in the dihydrolipoamide acetyltransferase component of the pyruvate dehydrogenase complex of Escherichia coli.
Related PDB
Related UniProtKB
[19]
Resource
Comments STRUCTURE BY NMR OF 1-78.
Medline ID 94222112
PubMed ID 8068086
Journal Eur J Biochem
Year 1994
Volume 221
Pages 87-100
Authors Berg A, de Kok A, Vervoort J
Title Sequential 1H and 15N nuclear magnetic resonance assignments and secondary structure of the N-terminal lipoyl domain of the dihydrolipoyl transacetylase component of the pyruvate dehydrogenase complex from Azotobacter vinelandii.
Related PDB
Related UniProtKB P10802
[20]
Resource
Comments
Medline ID
PubMed ID 7703242
Journal Biochemistry
Year 1995
Volume 34
Pages 4287-98
Authors Hendle J, Mattevi A, Westphal AH, Spee J, de Kok A, Teplyakov A, Hol WG
Title Crystallographic and enzymatic investigations on the role of Ser558, His610, and Asn614 in the catalytic mechanism of Azotobacter vinelandii dihydrolipoamide acetyltransferase (E2p).
Related PDB
Related UniProtKB
[21]
Resource
Comments
Medline ID
PubMed ID 8918601
Journal J Mol Biol
Year 1996
Volume 263
Pages 463-74
Authors Wallis NG, Allen MD, Broadhurst RW, Lessard IA, Perham RN
Title Recognition of a surface loop of the lipoyl domain underlies substrate channelling in the pyruvate dehydrogenase multienzyme complex.
Related PDB
Related UniProtKB
[22]
Resource
Comments STRUCTURE BY NMR OF 1-78.
Medline ID 97234563
PubMed ID 9119000
Journal Eur J Biochem
Year 1997
Volume 244
Pages 352-60
Authors Berg A, Vervoort J, de Kok A
Title Three-dimensional structure in solution of the N-terminal lipoyl domain of the pyruvate dehydrogenase complex from Azotobacter vinelandii.
Related PDB 1iyu 1iyv
Related UniProtKB P10802
[23]
Resource
Comments
Medline ID
PubMed ID 9280309
Journal FEBS Lett
Year 1997
Volume 413
Pages 339-43
Authors Allen MD, Perham RN
Title The catalytic domain of dihydrolipoyl acetyltransferase from the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus. Expression, purification and reversible denaturation.
Related PDB
Related UniProtKB
[24]
Resource
Comments
Medline ID
PubMed ID 9729480
Journal Biochem J
Year 1998
Volume 334
Pages 703-11
Authors Jackson JC, Vinluan CC, Dragland CJ, Sundararajan V, Yan B, Gounarides JS, Nirmala NR, Topiol S, Ramage P, Blume JE, Aicher TD, Bell PA, Mann WR
Title Heterologously expressed inner lipoyl domain of dihydrolipoyl acetyltransferase inhibits ATP-dependent inactivation of pyruvate dehydrogenase complex. Identification of important amino acid residues.
Related PDB
Related UniProtKB
[25]
Resource
Comments
Medline ID
PubMed ID 10419491
Journal J Biol Chem
Year 1999
Volume 274
Pages 21769-75
Authors Thelen JJ, Muszynski MG, David NR, Luethy MH, Elthon TE, Miernyk JA, Randall DD
Title The dihydrolipoamide S-acetyltransferase subunit of the mitochondrial pyruvate dehydrogenase complex from maize contains a single lipoyl domain.
Related PDB
Related UniProtKB
[26]
Resource
Comments
Medline ID
PubMed ID 10653630
Journal Biochemistry
Year 2000
Volume 39
Pages 872-9
Authors Spector S, Wang M, Carp SA, Robblee J, Hendsch ZS, Fairman R, Tidor B, Raleigh DP
Title Rational modification of protein stability by the mutation of charged surface residues.
Related PDB
Related UniProtKB
[27]
Resource
Comments
Medline ID
PubMed ID 10735853
Journal J Bacteriol
Year 2000
Volume 182
Pages 2119-24
Authors Stein A, Firshein W
Title Probable identification of a membrane-associated repressor of Bacillus subtilis DNA replication as the E2 subunit of the pyruvate dehydrogenase complex.
Related PDB
Related UniProtKB
[28]
Resource
Comments
Medline ID
PubMed ID 10913250
Journal Biochemistry
Year 2000
Volume 39
Pages 8448-59
Authors Jones DD, Stott KM, Howard MJ, Perham RN
Title Restricted motion of the lipoyl-lysine swinging arm in the pyruvate dehydrogenase complex of Escherichia coli.
Related PDB 1qjo
Related UniProtKB
[29]
Resource
Comments
Medline ID
PubMed ID 11368334
Journal Arch Biochem Biophys
Year 2001
Volume 386
Pages 123-35
Authors Liu S, Gong X, Yan X, Peng T, Baker JC, Li L, Robben PM, Ravindran S, Andersson LA, Cole AB, Roche TE
Title Reaction mechanism for mammalian pyruvate dehydrogenase using natural lipoyl domain substrates.
Related PDB
Related UniProtKB
[30]
Resource
Comments
Medline ID
PubMed ID 11114246
Journal J Mol Biol
Year 2001
Volume 305
Pages 49-60
Authors Jones DD, Stott KM, Reche PA, Perham RN
Title Recognition of the lipoyl domain is the ultimate determinant of substrate channelling in the pyruvate dehydrogenase multienzyme complex.
Related PDB
Related UniProtKB
[31]
Resource
Comments
Medline ID
PubMed ID 12173931
Journal Biochemistry
Year 2002
Volume 41
Pages 10446-53
Authors Jung HI, Cooper A, Perham RN
Title Identification of key amino acid residues in the assembly of enzymes into the pyruvate dehydrogenase complex of Bacillus stearothermophilus: a kinetic and thermodynamic analysis.
Related PDB
Related UniProtKB
[32]
Resource
Comments
Medline ID
PubMed ID 12526798
Journal Cell
Year 2003
Volume 112
Pages 113-22
Authors Jogl G, Tong L
Title Crystal structure of carnitine acetyltransferase and implications for the catalytic mechanism and fatty acid transport.
Related PDB
Related UniProtKB

Comments
This enzyme is Dihydrolipoyllysine-residue acetyltransferase, E2 component of pyruvate dehydrogenase complex.
The pyruvate dehydrogenase complex is composed of pyruvate dehydrogenaes (E1 component; E.C. 1.2.4.1), dihydrolipoyllysine S-acetyltransferase (E2 component; E.C. 2.3.1.12), and lipoamide dehydrogenase (E3 component; E.C. 1.8.1.4). (The E3 component corresponds to the entry T00017 in EzCatDB.)
This enzyme is composed of three N-terminal lipoyl-binding domains, E1/E3-binding domain, and the C-terminal catalytic domain. Although this enzyme has the same domain composition as that of its homologue (M00191 in EzCatDB), the catalytic residues are slightly different from those of the homologue. (The structure of the catalytic domain of this enzyme has not been solved yet.)
This enzyme catalyzes transfer of acetyl group from lypoyllysine of the lipoyl-binding domain to thiol group of CoA.
According to the literature [20], the reaction proceeds as follows:
(1) His610 acts as a general base to abstract a proton from the acceptor group, thiol group of CoA.
(2) Asn614 modulates the catalytic histidine, His610, along with the mainchain carbonyl oxygen of His375.
(3) The activated thiolate makes a nucleophilic attack on the transferred group, carbonyl carbon atom of the acetylated lipoamide substrate, resulting in the formation of a tetrahedral intermediate.
(4) This intermediate is stabilized by the hydroxyl group of Ser558' from the next subunit.
(5) The breakdown of this intermediate results in the transfer of the succinyl group to CoA and protonation of the leaving dithiolane group. Here, His610 acts as a general acid to protonate the leaving group.

Created Updated
2002-12-01 2009-02-26