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Literature summary for 2.7.1.180 extracted from

  • Deka, R.K.; Brautigam, C.A.; Liu, W.Z.; Tomchick, D.R.; Norgard, M.V.
    Molecular insights into the enzymatic diversity of flavin-trafficking protein (Ftp; formerly ApbE) in flavoprotein biogenesis in the bacterial periplasm (2016), MicrobiologyOpen, 5, 21-38 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.1.180

Crystallization (Commentary)

Crystallization (Comment) Organism
analysis of X-ray crystal structure of the Vibrio cholerae Nqr complex and structures of several purified components of the complex, PDB IDs 4U9S and 4P6V Vibrio cholerae serotype O1
purified Ftp_Ec mutant variant Y60N complexed with ADP, hanging drop vapor diffusion method, mixing of 0.004 ml of 20 mg/ml protein in Tris, pH 7.5, and 20 mM NaCl, with 0.04 ml of reservoir solution containing 0.2 M NH4NO3, and 20% w/v PEG 3350, and 0.001 ml of 50 mM MgCl2, and 0.001 ml of 50 mM ADP, 20°C, X-ray diffraction structure determination and analysis at 1.85 A resolution, molecular replacement and modelling Escherichia coli
purified recombinant wild-type enzyme Ftp_Ec, mutant E169K, and mutant Y60N bound to ADP, hanging drop vapor diffusion method, mixing of 0.004 ml of 20 mg/ml protein in 20 mM Tris, pH 7.5, and 20 mM NaCl, with 0.004 ml of reservoir solution containing 0.2 M NH4NO3 and 20% w/v PEG 3350 for the wild-type, and 25% w/v PEG 1500, 0.1 M MIB (malonate, imidazole and boric acid), pH 5.0 for the mutant, 20°C, X-ray diffraction structure determination and analysis at 1.75-1.88 A resolution Escherichia coli

Protein Variants

Protein Variants Comment Organism
E169K site-directed mutagenesis of the probabale catalytic site residue, the Ftp_EcE169K protein variant does not show binding of FAD, inactive mutant Escherichia coli
E169K site-directed mutagenesis, mutation of the active-site residue results in loss of FAD binding capability Escherichia coli
K207A site-directed mutagenesis, the mutant exhibits no detectable in vitro flavinylation activity Escherichia coli
Y60A site-directed mutagenesis, the engineered protein variant (Ftp_EcY60A) shows Mg2+-dependent FAD diphosphatase activity, but also retains its Mg2+-dependent FMN transferase (EC 2.7.1.180) activity on the protein substrate, indicating that the protein variant enzyme has dual activity Escherichia coli
Y60N site-directed mutagenesis, a single amino acid substitution converts it from an FAD-binding protein to a Mg2+-dependent FAD diphosphatase (Ftp_Tp-like) Escherichia coli
Y60N site-directed mutagenesis, the single amino acid substitution converts it from an FAD-binding protein to a Mg2+-dependent FAD pdiphosphatase (Ftp_Tp-like, EC 3.6.1.18). The Ftp_EcY60A protein variant binds FAD, rapidly hydrolyzes it, and the product FMN dissociates. But the mutant also retains its Mg2+-dependent FMN transferase (EC 2.7.1.180) activity on the protein substrate. As the site of attack for the FMN transferase reaction is the beta-phosphate of the FAD, and given the large distance between the two metals in the ADP-inhibited Ftp_EcY60N structure, it is reasonable to expect that only metal site 2 requires a Mg2+ ion for this activity Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
ADP the Ftp_EcY60N enzyme mutant displays FAD diphosphatase activity that is inhibited by ADP Escherichia coli
EDTA
-
 Escherichia coli
EDTA
-
 Treponema pallidum
EDTA
-
 Vibrio cholerae serotype O1

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information single-turnover kinetics Escherichia coli

Localization

Localization Comment Organism GeneOntology No. Textmining
periplasm
-
 Treponema pallidum
-
-
periplasm
-
 Escherichia coli
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ presence of a Ca2+ ion in metal site 1 in the apo form of mutant Ftp_EcY60N, the Ca2+ ion serves a role in properly positioning substrate and protein residues Escherichia coli
Mg2+ dependent on, metal-dependent enzyme Treponema pallidum
Mg2+ dependent on, metal-dependent enzyme Vibrio cholerae serotype O1
Mg2+ dependent on, metal-dependent enzyme, bimetal center in the crystal structure of Escherichia coli Ftp Escherichia coli
Mg2+ required, dependent on, Mg2+ ion is directly involved in catalysis. A single metal ion is bound in the wild-type enzyme and mutant Ftp_EcE169K structures. The inhibited mutant Ftp_EcY60N contains a bimetal Mg2+ center Escherichia coli
additional information identification of a bimetal center in the crystal structure of Escherichia coli Ftp (Ftp_Ec) Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
FAD + L-threonyl-[protein] Treponema pallidum
-
 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein] Escherichia coli
-
 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein] Vibrio cholerae serotype O1
-
 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein] Vibrio cholerae serotype O1 Classical Ogawa 395
-
 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein] Vibrio cholerae serotype O1 ATCC 39541
-
 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein] Treponema pallidum Nichols
-
 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein] Vibrio cholerae serotype O1 O395
-
 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + [protein]-L-threonine Escherichia coli
-
 [protein]-FMN-L-threonine + AMP
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P0AB85
-
-
Treponema pallidum O83774
-
-
Treponema pallidum Nichols O83774
-
-
Vibrio cholerae serotype O1 A5F5Y3
-
-
Vibrio cholerae serotype O1 ATCC 39541 A5F5Y3
-
-
Vibrio cholerae serotype O1 Classical Ogawa 395 A5F5Y3
-
-
Vibrio cholerae serotype O1 O395 A5F5Y3
-
-

Source Tissue

Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
FAD + L-threonyl-[NqrC protein]
-
 Treponema pallidum AMP + FMN-L-threonyl-[NqrC protein] + H+
-
 ?
FAD + L-threonyl-[NqrC protein]
-
 Escherichia coli AMP + FMN-L-threonyl-[NqrC protein] + H+
-
 ?
FAD + L-threonyl-[NqrC protein]
-
 Vibrio cholerae serotype O1 AMP + FMN-L-threonyl-[NqrC protein] + H+
-
 ?
FAD + L-threonyl-[NqrC protein]
-
 Vibrio cholerae serotype O1 Classical Ogawa 395 AMP + FMN-L-threonyl-[NqrC protein] + H+
-
 ?
FAD + L-threonyl-[NqrC protein]
-
 Vibrio cholerae serotype O1 ATCC 39541 AMP + FMN-L-threonyl-[NqrC protein] + H+
-
 ?
FAD + L-threonyl-[NqrC protein]
-
 Vibrio cholerae serotype O1 O395 AMP + FMN-L-threonyl-[NqrC protein] + H+
-
 ?
FAD + L-threonyl-[protein]
-
 Treponema pallidum AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein]
-
 Escherichia coli AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein]
-
 Vibrio cholerae serotype O1 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein] RnfG_Ec subunit of the Rnf_Ec-redox system of Escherichia coli Treponema pallidum AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein]
-
 Vibrio cholerae serotype O1 Classical Ogawa 395 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein]
-
 Vibrio cholerae serotype O1 ATCC 39541 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein]
-
 Treponema pallidum Nichols AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein] RnfG_Ec subunit of the Rnf_Ec-redox system of Escherichia coli Treponema pallidum Nichols AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[protein]
-
 Vibrio cholerae serotype O1 O395 AMP + FMN-L-threonyl-[protein] + H+
-
 ?
FAD + L-threonyl-[RnfG protein] RnfG subunit of the Rnf redox system Treponema pallidum AMP + FMN-L-threonyl-[RnfG protein] + H+
-
 ?
FAD + L-threonyl-[RnfG protein] RnfG subunit of the Rnf redox system Escherichia coli AMP + FMN-L-threonyl-[RnfG protein] + H+
-
 ?
FAD + L-threonyl-[RnfG protein] RnfG subunit of the Rnf redox system Vibrio cholerae serotype O1 AMP + FMN-L-threonyl-[RnfG protein] + H+
-
 ?
FAD + L-threonyl-[RnfG protein] RnfG subunit of the Rnf redox system Vibrio cholerae serotype O1 Classical Ogawa 395 AMP + FMN-L-threonyl-[RnfG protein] + H+
-
 ?
FAD + L-threonyl-[RnfG protein] RnfG subunit of the Rnf redox system Vibrio cholerae serotype O1 ATCC 39541 AMP + FMN-L-threonyl-[RnfG protein] + H+
-
 ?
FAD + L-threonyl-[RnfG protein] RnfG subunit of the Rnf redox system Vibrio cholerae serotype O1 O395 AMP + FMN-L-threonyl-[RnfG protein] + H+
-
 ?
FAD + L-threonyl-[Shewanella oneidensis NqrC protein] a high-resolution structure of the Ftp-mediated flavinylated protein of Shewanella oneidensis NqrC identifies an essential lysine in phosphoester-threonyl-FMN bond formation in the posttranslationally modified flavoproteins Escherichia coli AMP + FMN-L-threonyl-[Shewanella oneidensis NqrC protein] + H+
-
 ?
FAD + [periplasmic redox-carrying protein RnfG_Ec]-L-threonine flavinylating via the metal-dependent covalent attachment of FMN Escherichia coli [periplasmic redox-carrying protein RnfG_Ec]-FMN-L-threonine + AMP
-
 ?
FAD + [protein]-L-threonine
-
 Escherichia coli [protein]-FMN-L-threonine + AMP
-
 ?
FAD + [Shewanella oneidensis protein NqrC]-L-threonine a subunit (NqrC) of a cytoplasmic membrane redox system (Nqr), detection of an essential lysine residue in phosphoester-threonyl-FMN bond formation in the posttranslationally modified flavoprotein Escherichia coli [Shewanella oneidensis protein NqrC]-FMN-L-threonine + AMP
-
 ?
additional information the flavin-trafficking protein (Ftp) catalyzes the transfer of the FMN moiety of FAD and its covalent binding to the hydroxyl group of a threonine residue in a target flavoprotein (EC 2.7.1.180). The enzyme is capable of flavinylating periplasmic redox-carrying proteins (e.g., RnfG_Ec) via the metal-dependent covalent attachment of FMN. A single amino acid substitution Y60N converts it from an FAD-binding protein to a Mg2+-dependent FAD diphosphatase (Ftp_Tp-like) (EC 3.6.1.18). The engineered protein variant (Ftp_EcY60A) shows Mg2+-dependent FAD diphosphatase activity, but also retains its Mg2+-dependent FMN transferase (EC 2.7.1.180) activity on the protein substrate, indicating that the protein variant enzyme has dual activity. The Ftp_EcY60A protein variant binds FAD, yet rapidly hydrolyzes it and the product FMN dissociates. Substrate binding structures, detailed overview Escherichia coli ?
-
-
additional information the flavin-trafficking protein (Ftp) catalyzes the transfer of the FMN moiety of FAD and its covalent binding to the hydroxyl group of a threonine residue in a target flavoprotein (EC 2.7.1.180). The enzyme is capable of flavinylating periplasmic redox-carrying proteins (e.g., RnfG_Ec) via the metal-dependent covalent attachment of FMN. It also displays FAD diphosphatase activity in vitro, hydrolyzing FAD into FMN and AMP (EC 3.6.1.18). Substrate binding structures, detailed overview Treponema pallidum ?
-
-
additional information the Escherichia coli enzyme shows no Mg2+-dependent FAD pyrophosphatase (EC 3.6.1.18) activity Escherichia coli ?
-
-
additional information the flavin-trafficking protein (Ftp) catalyzes the transfer of the FMN moiety of FAD and its covalent binding to the hydroxyl group of a threonine residue in a target flavoprotein (EC 2.7.1.180). The enzyme is capable of flavinylating periplasmic redox-carrying proteins (e.g., RnfG_Ec) via the metal-dependent covalent attachment of FMN. It also displays FAD diphosphatase activity in vitro, hydrolyzing FAD into FMN and AMP (EC 3.6.1.18). Substrate binding structures, detailed overview Treponema pallidum Nichols ?
-
-

Synonyms

Synonyms Comment Organism
AbpE
-
 Escherichia coli
apbE
-
 Treponema pallidum
apbE
-
 Escherichia coli
apbE
-
 Vibrio cholerae serotype O1
flavin-trafficking protein
-
 Escherichia coli
FMN transferase
-
 Treponema pallidum
FMN transferase
-
 Escherichia coli
FMN transferase
-
 Vibrio cholerae serotype O1
Ftp
-
 Treponema pallidum
Ftp
-
 Escherichia coli
Ftp
-
 Vibrio cholerae serotype O1
Ftp_Ec
-
 Escherichia coli
Mg2+-dependent FMN transferase
-
 Treponema pallidum
Mg2+-dependent FMN transferase
-
 Escherichia coli
Mg2+-dependent FMN transferase
-
 Vibrio cholerae serotype O1
More cf. EC 3.6.1.18 Treponema pallidum
More cf. EC 3.6.1.18 Escherichia coli
More cf. EC 3.6.1.18 Vibrio cholerae serotype O1
protein-dependent FMN transferase
-
 Treponema pallidum
protein-dependent FMN transferase
-
 Escherichia coli
protein-dependent FMN transferase
-
 Vibrio cholerae serotype O1

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Escherichia coli
37
-
 assay at Treponema pallidum
37
-
 assay at Escherichia coli
37
-
 assay at Vibrio cholerae serotype O1

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Treponema pallidum
7.5
-
 assay at Escherichia coli
7.5
-
 assay at Vibrio cholerae serotype O1

General Information

General Information Comment Organism
evolution there likely are two classes of Ftps, one associated with FAD-binding and the other with FAD hydrolysis Treponema pallidum
evolution there likely are two classes of Ftps, one associated with FAD-binding and the other with FAD hydrolysis Escherichia coli
evolution there likely are two classes of Ftps, one associated with FAD-binding and the other with FAD hydrolysis Vibrio cholerae serotype O1
malfunction a single amino acid substitution Y60N converts it from an FAD-binding protein to a Mg2+-dependent FAD diphosphatase (Ftp_Tp-like). The engineered protein variant (Ftp_EcY60A) shows Mg2+-dependent FAD diphosphatase activity, but also retains its Mg2+-dependent FMN transferase (EC 2.7.1.180) activity on the protein substrate, indicating that the protein variant enzyme has dual activity Escherichia coli
malfunction a single amino acid substitution converts it from an FAD-binding protein to a Mg2+-dependent FAD diphosphatase (Ftp_Tp-like, EC 3.6.1.18) Escherichia coli
additional information the critical residue that contacts the isoalloxazine ring of FAD, is a tyrosine residue in the FAD-binding Ftps Treponema pallidum
additional information the critical residue that contacts the isoalloxazine ring of FAD, is a tyrosine residue in the FAD-binding Ftps. FAD binding structure involving residue K207, overview Escherichia coli
physiological function the flavin-trafficking protein (Ftp) catalyzes the transfer of the FMN moiety of FAD to a threonine residue in a target flavoprotein. Both types of Ftps are capable of flavinylating periplasmic redox-carrying proteins (e.g., RnfG_Ec) via the metal-dependent covalent attachment of FMN. Possible mechanism by which flavoproteins are generated, overview Escherichia coli
physiological function the flavin-trafficking protein (Ftp) catalyzes the transfer of the FMN moiety of FAD to a threonine residue in a target flavoprotein. Both types of Ftps are capable of flavinylating periplasmic redox-carrying proteins (e.g., RnfG_Ec) via the metal-dependent covalent attachment of FMN. Possible mechanism by which flavoproteins are generated, overview Vibrio cholerae serotype O1
physiological function the flavin-trafficking protein (Ftp) in the syphillis spirochete Treponema pallidum (Ftp_Tp) is a bacterial metal-dependent FAD diphosphatase that hydrolyzes FAD into AMP and FMN in the periplasm. Both types of Ftps are capable of flavinylating periplasmic redox-carrying proteins (e.g., RnfG_Ec) via the metal-dependent covalent attachment of FMN. Possible mechanism by which flavoproteins are generated, overview Treponema pallidum
physiological function the enzyme flavinylate the redox subunit, NqrC, via its metal-dependent FMN transferase activity Escherichia coli