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Literature summary for 4.1.99.26 extracted from

  • Khaliullin, B.; Ayikpoe, R.; Tuttle, M.; Latham, J.
    Mechanistic elucidation of the mycofactocin-biosynthetic radical S-adenosylmethionine protein, MftC (2017), J. Biol. Chem., 292, 13022-13033 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.1.99.26

Activating Compound

Activating Compound Comment Organism Structure
MftB required for activity Mycobacterium ulcerans

Cloned(Commentary)

Cloned (Comment) Organism
gene mftC, recombinant expression of His-tagged enzyme from plasmid pPH151, which contains the isc operon, in Escherichia coli strain Bl21(DE3) Mycobacterium ulcerans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + S-adenosyl-L-methionine + AH2 Mycobacterium ulcerans
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 C-terminal [mycofactocin precursor peptide]-glycyl-3-amino-5-[(4-hydroxyphenyl)methyl]-4,4-dimethylpyrrolidin-2-one + 5'-deoxyadenosine + L-methionine + A
-
 ?
C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + S-adenosyl-L-methionine + AH2 Mycobacterium ulcerans Agy99
-
 C-terminal [mycofactocin precursor peptide]-glycyl-3-amino-5-[(4-hydroxyphenyl)methyl]-4,4-dimethylpyrrolidin-2-one + 5'-deoxyadenosine + L-methionine + A
-
 ?

Organism

Organism UniProt Comment Textmining
Mycobacterium ulcerans A0PM49
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Mycobacterium ulcerans A0PM49 cf. EC 1.3.98.7
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Mycobacterium ulcerans Agy99 A0PM49
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Mycobacterium ulcerans Agy99 A0PM49 cf. EC 1.3.98.7
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Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration Mycobacterium ulcerans

Reaction

Reaction Comment Organism Reaction ID
C-terminal [mycofactocin precursor peptide]-glycyl-3-amino-5-[(4-hydroxyphenyl)methyl]-4,4-dimethylpyrrolidin-2-one + 5'-deoxyadenosine + L-methionine + A = C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + S-adenosyl-L-methionine + AH2 enzyme mechanism, detailed overview. MftC catalyzes the formation of two products from substrate MftA, 13C NMR and 1H NMR analysis of substrate, intermediates and products. The major product of the MftC reaction is a Val-Tyr* crosslinked peptide formed through two S-adenosylmethionine-dependent turnovers. The hydroxyl group on MftA Tyr30 is required for MftC catalysis Mycobacterium ulcerans
a

Renatured (Commentary)

Renatured (Comment) Organism
reconstitution of purified recombinant His-tagged enzyme MftC by addition of DTT and FeCl3, followed by Na2S addition, and ultrafiltration Mycobacterium ulcerans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + S-adenosyl-L-methionine + AH2
-
 Mycobacterium ulcerans C-terminal [mycofactocin precursor peptide]-glycyl-3-amino-5-[(4-hydroxyphenyl)methyl]-4,4-dimethylpyrrolidin-2-one + 5'-deoxyadenosine + L-methionine + A
-
 ?
C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + S-adenosyl-L-methionine + AH2 enzyme MftC catalyzes the oxidative decarboxylation of the C-terminal tyrosine (Tyr30) on the mycofactocin precursor peptide MftA. MftC catalyzes the formation of two MftA products annotated as MftA* and MftA**. The MftAM1W variant is used to increase the spectroscopic handle at 280 nm. The major product, MftA*, is a tyramine-valine-cross-linked peptide formed by MftC through two S-adenosylmethionine-dependent turnovers. In case of the reaction containing MftA M1W/V29A, HPLC analysis shows a a minor peak for MftA M1W/V29A* (10%) and a major peak for MftA M1W/V29A**. Substitution in the penultimate MftA Val29 position causes the accumulation of an MftA** minor product. The 1H-NMR spectrum indicates that this minor product contains an alpha/beta-unsaturated bond that likely arises from an aborted intermediate of MftA* synthesis Mycobacterium ulcerans C-terminal [mycofactocin precursor peptide]-glycyl-3-amino-5-[(4-hydroxyphenyl)methyl]-4,4-dimethylpyrrolidin-2-one + 5'-deoxyadenosine + L-methionine + A
-
 ?
C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + S-adenosyl-L-methionine + AH2
-
 Mycobacterium ulcerans Agy99 C-terminal [mycofactocin precursor peptide]-glycyl-3-amino-5-[(4-hydroxyphenyl)methyl]-4,4-dimethylpyrrolidin-2-one + 5'-deoxyadenosine + L-methionine + A
-
 ?
C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + S-adenosyl-L-methionine + AH2 enzyme MftC catalyzes the oxidative decarboxylation of the C-terminal tyrosine (Tyr30) on the mycofactocin precursor peptide MftA. MftC catalyzes the formation of two MftA products annotated as MftA* and MftA**. The MftAM1W variant is used to increase the spectroscopic handle at 280 nm. The major product, MftA*, is a tyramine-valine-cross-linked peptide formed by MftC through two S-adenosylmethionine-dependent turnovers. In case of the reaction containing MftA M1W/V29A, HPLC analysis shows a a minor peak for MftA M1W/V29A* (10%) and a major peak for MftA M1W/V29A**. Substitution in the penultimate MftA Val29 position causes the accumulation of an MftA** minor product. The 1H-NMR spectrum indicates that this minor product contains an alpha/beta-unsaturated bond that likely arises from an aborted intermediate of MftA* synthesis Mycobacterium ulcerans Agy99 C-terminal [mycofactocin precursor peptide]-glycyl-3-amino-5-[(4-hydroxyphenyl)methyl]-4,4-dimethylpyrrolidin-2-one + 5'-deoxyadenosine + L-methionine + A
-
 ?
additional information site-directed mutagenesis of MftA Tyr30 indicates that a labile proton is required for catalysis. SUbstrate mutants used are MftA M1W, MftA M1W/Y30F, MftA M1W/Y30S, and MftA M1W/Y30W Mycobacterium ulcerans ?
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additional information site-directed mutagenesis of MftA Tyr30 indicates that a labile proton is required for catalysis. SUbstrate mutants used are MftA M1W, MftA M1W/Y30F, MftA M1W/Y30S, and MftA M1W/Y30W Mycobacterium ulcerans Agy99 ?
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Synonyms

Synonyms Comment Organism
mftC
-
 Mycobacterium ulcerans
mycofactocin maturase
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 Mycobacterium ulcerans
mycofactocin-biosynthetic radical S-adenosylmethionine protein
-
 Mycobacterium ulcerans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at room temperature Mycobacterium ulcerans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Mycobacterium ulcerans

Cofactor

Cofactor Comment Organism Structure
S-adenosyl-L-methionine MftC utilizes 2 eq of SAM to crosslink Tyr30 and Val29 Mycobacterium ulcerans

General Information

General Information Comment Organism
evolution MftC belongs to a subfamily of RS proteins that contain an about 100-amino acid C-terminal domain annotated as a SPASM domain, named after subtilosin A, pyrroloquinoline quinone, anaerobic sulfatase maturating enzyme, and mycofactocin Mycobacterium ulcerans
physiological function MftC catalyzes the formation of two isomeric products. The major product, MftA*, is a tyramine-valine-cross-linked peptide formed by MftC through two S-adenosylmethionine-dependent turnovers. The hydroxyl group on MftA Tyr30 is required for MftC catalysis. A substitution in the penultimate substrate MftA Val29 position causes the accumulation of an MftA** minor product which contains an alphabeta-unsaturated bond that likely arises from an aborted intermediate of MftA* synthesis Mycobacterium ulcerans
physiological function the enzyme substrate MftA is about 30-60 amino acids in length and contains a conserved C-terminal sequence (-IDGXCGVY). The C-terminus is putatively modified by the remaining gene products to form mycofactocin, which is anticipated to serve as a redox cofactor for other nicotinamide-dependent proteins. MftC is a radical S-adenosyl-L-methionine (RS) protein responsible for the first step of the biosynthesis of mycofactocin. MftC catalyzes the formation of two isomeric MftA products annotated as MftA* and MftA**. And MftC catalyzes the conversion of MftA** to MftA*. MftC catalyzes a diverse array of chemical transformations, catalytic mechanism, overview Mycobacterium ulcerans