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Literature summary extracted from

  • Schneider, K.; Dimroth, P.; Bott, M.
    Biosynthesis of the prosthetic group of citrate lyase (2000), Biochemistry, 39, 9438-9450.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.52 expressed in Escherichia coli Escherichia coli
2.7.7.61 expressed in Escherichia coli BL21-DE3 Escherichia coli
4.1.3.6 apo-ACP Klebsiella pneumoniae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.52 32600
-
 ? * 32600, SDS-PAGE Escherichia coli
2.7.7.61 20300
-
 x * 20300, SDS-PAGE, autoradiography Escherichia coli
4.1.3.6 11760
-
 apo-ACP-His, mass spectrometry Escherichia coli
4.1.3.6 11760
-
 apo-ACP-His, mass spectrometry Klebsiella pneumoniae
4.1.3.6 12500
-
 apo-ACP-His, SDS-PAGE Escherichia coli
4.1.3.6 12500
-
 apo-ACP-His, SDS-PAGE Klebsiella pneumoniae
4.1.3.6 12650
-
 holo-ACP-His, mass spectrometry Escherichia coli
4.1.3.6 12650
-
 holo-ACP-His, mass spectrometry Klebsiella pneumoniae
4.1.3.6 15000
-
 holo-ACP-His, SDS-PAGE Escherichia coli
4.1.3.6 15000
-
 holo-ACP-His, SDS-PAGE Klebsiella pneumoniae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.52 ATP + 3'-dephospho-CoA Escherichia coli
-
 2'-(5''-triphosphoribosyl)-3'-dephospho-CoA + adenine
-
 ?
4.1.3.6 citrate Escherichia coli
-
 acetate + oxaloacetate
-
 ?
4.1.3.6 citrate Klebsiella pneumoniae
-
 acetate + oxaloacetate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.52 Escherichia coli
-
 strain JM83, strain CM2080, gene citG
-
2.4.2.52 Escherichia coli P77231
-
-
2.4.2.52 Klebsiella pneumoniae P45414
-
-
2.7.7.61 Escherichia coli
-
-
-
4.1.3.6 Escherichia coli
-
-
-
4.1.3.6 Klebsiella pneumoniae
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.52
-
 Escherichia coli
2.7.7.61
-
 Escherichia coli
4.1.3.6 apo-ACP Klebsiella pneumoniae
4.1.3.6 apo-ACP, i.e. gamma-subunit Escherichia coli

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.7.61 2'-(5-triphosphoribosyl)-3'-dephospho-CoA + apo-[citrate (pro-3S)-lyase] = diphosphate + holo-[citrate (pro-3S)-lyase] 2'-(5"-triphosphoribosyl)-3'-dephospho-CoA: prosthetic group of the gamma-subunit of the citrate lyase Escherichia coli
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.1.3.6 8
-
-
 Klebsiella pneumoniae

Storage Stability

EC Number Storage Stability Organism
2.4.2.52 -4°C Escherichia coli
2.7.7.61 4°C Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.52 ATP + 3'-dephospho-CoA
-
 Escherichia coli 2'-(5''-triphosphoribosyl)-3'-dephospho-CoA + adenine
-
 ?
2.4.2.52 ATP + 3'-dephospho-CoA ATP cannot be replaced by CTP, GTP or UTP Escherichia coli 2'-(5''-triphosphoribosyl)-3'-dephospho-CoA + adenine
-
 ?
2.4.2.52 additional information holo-acyl carrier protein formation proceeds as follows. First, a prosthetic group precursor, presumably 2'-(5''-triphosphoribosyl)-3'-dephospho-CoA, is formed from ATP and dephospho-CoA in a reaction catalyzed by CitG. Second, holo-ACP is formed from apo-ACP and the prosthetic group precursor in a reaction catalyzed by CitX. The conversion of apo-cyl carrier protein into holo-cyl carrier protein is achieved in vitro by incubation of apo-cyl carrier protein with proteins CitX, CitG, ATP, and dephospho-CoA. ATP cannot be substituted with GTP, CTP, UTP, ADP, or AMP. In the absence of CitG or dephospho-CoA, AMP-cyl carrier protein is formed. It is not possible to further convert AMP-cyl carrier protein to holo-cyl carrier protein by subsequent incubation with CitG and dephospho-CoA Klebsiella pneumoniae ?
-
 ?
2.4.2.52 additional information holo-acyl carrier protein formation proceeds as follows. First, a prosthetic group precursor, presumably 2'-(5''-triphosphoribosyl)-3'-dephospho-CoA, is formed from ATP and dephospho-CoA in a reaction catalyzed by CitG. Second, holo-ACP is formed from apo-ACP and the prosthetic group precursor in a reaction catalyzed by CitX. The conversion of apo-cyl carrier protein into holo-cyl carrier protein is achieved in vitro by incubation of apo-cyl carrier protein with proteins CitX, CitG, ATP, and dephospho-CoA. ATP cannot be substituted with GTP, CTP, UTP, ADP, or AMP. In the absence of CitG or dephospho-CoA, AMP-cyl carrier protein is formed. It is not possible to further convert AMP-cyl carrier protein to holo-cyl carrier protein by subsequent incubation with CitG and dephospho-CoA Escherichia coli ?
-
 ?
2.7.7.61 apo-ACP + ATP side activity of the CitX in absence of its natural substrates, besides ATP, CTP, GTP, and UTP serve as nucleotidyl donors in vitro, ACP: acyl carrier protein, gamma-subunit of the citrate lyase Escherichia coli AMP-ACP + diphosphate
-
 ?
2.7.7.61 additional information no substrate in vitro: NAD+ Escherichia coli ?
-
 ?
4.1.3.6 citrate
-
 Escherichia coli acetate + oxaloacetate
-
 ?
4.1.3.6 citrate
-
 Klebsiella pneumoniae acetate + oxaloacetate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.2.52 ? ? * 32600, SDS-PAGE Escherichia coli
2.7.7.61 ? x * 20300, SDS-PAGE, autoradiography Escherichia coli
4.1.3.6 ? alpha, beta, gamma Klebsiella pneumoniae

Synonyms

EC Number Synonyms Comment Organism
2.4.2.52 CitG
-
 Klebsiella pneumoniae
2.4.2.52 CitG
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.3.6 25
-
 assay at Klebsiella pneumoniae

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.3.6 2'-(5''-phosphoribosyl)-3'-dephospho-coenzyme A attached to the gamma subunit, i.e. apo-ACP, via a phosphodiester linkage to serine-14, CitG and CitX catalyze the initial formation of the precursor 2'-(5''-triphosphoribosyl)-3'-dephospho-CoA and subsequent transfer to apo-citrate lyase with removal of diphosphate Klebsiella pneumoniae
4.1.3.6 2'-(5''-phosphoribosyl)-3'-dephospho-coenzyme A attached to the gamma subunit, i.e. apo-ACP, via phosphodiester linkage by CitX, resulting in the formation of holo-ACP Escherichia coli