Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 4.1.99.14 extracted from

  • Yang, L.; Lin, G.; Nelson, R.S.; Jian, Y.; Telser, J.; Li, L.
    Mechanistic studies of the spore photoproduct lyase via a single cysteine mutation (2012), Biochemistry, 51, 7173-7188.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene splB, overexpression of His6-tagged wild-type and mutant enzymes and co-expression with plasmid pDB1282, in Escherichia coli strain BL21(DE3) Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
C141A site-directed mutagenesis, the mutant repairs spore photoproduct at a rate which is about 3fold slower than the wild-type enzyme, the mutant shows altered competitive kinetic isotope effects compared to the wild-type enzyme. S-adenosyl-L-methionine plays a non-catalytical role in the mutant in contrast to the wild-type enzyme Bacillus subtilis

Inhibitors

Inhibitors Comment Organism Structure
Dithionite
-
Bacillus subtilis
DTT
-
Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state kinetics and competitive kinetic isotope effects of recombinant wild-type and mutant enzymes, overview Bacillus subtilis

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ metalloenzyme, the enzyme uses a [4Fe-4S]1+ cluster to reduce the S-adenosyl-L-methionine generating a catalytic 5'-deoxyadenosyl radical Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine Bacillus subtilis
-
thymidylyl-(3'->5')-thymidylate
-
?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine Bacillus subtilis 168
-
thymidylyl-(3'->5')-thymidylate
-
?

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
gene splB
-
Bacillus subtilis 168
-
gene splB
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity and cation exchange chromatography Bacillus subtilis

Reaction

Reaction Comment Organism Reaction ID
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double-helical DNA) = thymidylyl-(3'->5')-thymidylate (in double-helical DNA) reaction mechanism, overview Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine
-
Bacillus subtilis thymidylyl-(3'->5')-thymidylate
-
?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine the reaction is highly stereo-selective Bacillus subtilis thymidylyl-(3'->5')-thymidylate
-
?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine
-
Bacillus subtilis 168 thymidylyl-(3'->5')-thymidylate
-
?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine the reaction is highly stereo-selective Bacillus subtilis 168 thymidylyl-(3'->5')-thymidylate
-
?

Synonyms

Synonyms Comment Organism
SPL
-
Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
assay at room temperature Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
S-adenosyl-L-methionine a radical S-adenosyl-L-methionine enzyme that uses a [4Fe-4S]1+ cluster to reduce the S-adenosyl-L-methionine generating a catalytic 5'-deoxyadenosyl radical Bacillus subtilis

pI Value

Organism Comment pI Value Maximum pI Value
Bacillus subtilis recombinant His6-tagged mutant C141A, isoelectric focussing
-
8.5

General Information

General Information Comment Organism
evolution the enzyme is a member of the radical SAM superfamily Bacillus subtilis
malfunction the enzyme C141A mutant produces thymidylyl-(3'->5')-thymidylate likely via an unprecedented thymine radical cation reduction (proton coupled electron transfer) mechanism Bacillus subtilis
additional information residue C141 is solvent exposable and no other protein residue locates between the thymidylyl-(3'->5')-thymidylate radical and the C141 in the wild--type enzyme reaction pathway Bacillus subtilis
physiological function spore photoproduct lyase repairs a covalent UV-induced thymine dimer, spore photoproduct, in germinating endospores and is responsible for endospores' strong UV resistance. The spore photoproduct is rapidly repaired by the metalloenzyme spore photoproduct lyase when spores start germinating Bacillus subtilis