Information on EC 1.3.98.7 - [mycofactocin precursor peptide]-tyrosine decarboxylase

mycofactocin maturase

5 entries

Rv0693

3 entries

C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-L-tyrosine + S-adenosyl-L-methionine = C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + CO2 + 5'-deoxyadenosine + L-methionine

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C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-L-tyrosine L-tyrosine-carboxylyase

This is a bifunctional radical AdoMet (radical SAM) enzyme that catalyses the first two steps in the biosynthesis of the enzyme cofactor mycofactocin. Activity requires the presence of the MftB chaperone. The other activity of the enzyme is EC 4.1.99.26, 3-amino-5-[(4-hydroxyphenyl)methyl]-4,4-dimethylpyrrolidin-2-one synthase.

C-terminal [mycofactocin precursor peptide MftA]-glycyl-L-valyl-L-tyrosine + S-adenosyl-L-methionine

C-terminal [mycofactocin precursor peptide MftA]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + CO2 + 5'-deoxyadenosine + L-methionine

C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-L-tyrosine + S-adenosyl-L-methionine

C-terminal [mycofactocin precursor peptide]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + CO2 + 5'-deoxyadenosine + L-methionine

additional information

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C-terminal [mycofactocin precursor peptide MftA]-glycyl-L-valyl-L-tyrosine + S-adenosyl-L-methionine

C-terminal [mycofactocin precursor peptide MftA]-glycyl-L-valyl-4-[2-aminoethenyl]phenol + CO2 + 5'-deoxyadenosine + L-methionine

S-adenosyl-L-methionine

residues Cys30, 34, and 37 are the radical S-adenosylmethionine ligands

[4Fe-4S]-center

MftC binds a radical S-adenosylmethionine [4Fe-4S] cluster and two auxiliary [4Fe-4S] clusters that are required for MftA modification. Presence of S-adenosylmethionine and MftA affects the environments of the radical S-adenosylmethionine and Aux I cluster whereas the Aux II cluster is unaffected by the substrates. All three cluster are required for cataylsis

Iron

Mycobacterium tuberculosis

A0QSB8

purified and reconstituted protein contains 10.8 mol of iron and 7.4 mol of sulfide per mol of MftC

760576

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760787

P9WJ79

UniProt

Mycobacterium tuberculosis H37Rv

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760787

P9WJ79

UniProt

Mycobacterium ulcerans Agy99

Mycolicibacterium smegmatis ATCC 700084

physiological function

7 entries

C251A

insoluble

C269A

auxiliary [4Fe-4S] cluster I mutant, capable of catalyzing the reductive cleavage of SAM to form dAdo but incapable of converting MftA to MftA* or MftA**

C269A/C323A

insoluble

C30A/C34A/C37A

insoluble

C30A/C37A

radical S-adenosylmethionine mutant, can neither cleave SAM nor modify MftA

C310A/C341A

auxiliary [4Fe-4S] cluster II mutant, capable of catalyzing the reductive cleavage of SAM to form dAdo, incapable of converting MftA to MftA* or MftA**

C323A

auxiliary [4Fe-4S] cluster I mutant, capable of catalyzing the reductive cleavage of SAM to form dAdo, incapable of converting MftA to MftA* or MftA**

additional information

systematical replacement of Cys residues by Ala. The RS KO could neither cleave SAM nor modify MftA, consistent with the successful knockout of the RS cluster. Activity assays for Aux I and Aux II KO's also provided insightful results. Both Aux I and Aux II KO's were capable of catalyzing the reductive cleavage of SAM to form dAdo (Figure 3A), suggesting that the RS cluster remained intact and in an active conformation in the mutated proteins. However, when assayed against MftA, both Aux I and Aux II KO's were incapable of converting MftA to MftA* or MftA**

recombinant protein, anaerobically purified using a N-terminal His6-tag. Enzyme is brown in color

A0QSB8

760576

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