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Literature summary for 2.8.1.4 extracted from

  • Liu, Y.; Zhu, X.; Nakamura, A.; Orlando, R.; Soell, D.; Whitman, W.B.
    Biosynthesis of 4-thiouridine in tRNA in the methanogenic archaeon Methanococcus maripaludis (2012), J. Biol. Chem., 287, 36683-36692.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.8.1.4

Cloned(Commentary)

Cloned (Comment) Organism
-
 Methanococcus maripaludis

Protein Variants

Protein Variants Comment Organism
C265A cells expressing the C265A variant show 88% decrease in their 4-thiouridine level. The mutant enzyme contains no detectable amount of persulfide Methanococcus maripaludis
C268A cells expressing the C265A variant contain no 4-thiouridine. The mutant enzyme contains no detectable amount of persulfide Methanococcus maripaludis
C348A cells expressing the C348A variant show 33% decreases in their 4-thiouridine levels. The mutant enzyme contains no detectable amount of persulfide Methanococcus maripaludis
C78A cells expressing the C78A variant show 8% decrease in their 4-thiouridine levels. The C78A variant has similar amounts of persulfide as the wild-type protein Methanococcus maripaludis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1
-
 Sulfide pH 7.3, 37°C Methanococcus maripaludis

Metals/Ions

Metals/Ions Comment Organism Structure
additional information no spectroscopic features indicative of Fe-S clusters are present Methanococcus maripaludis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
sulfide + tRNA-uridine Methanococcus maripaludis the enzyme is invoved in biosynthesis of 4-thiouridine in tRNA. 4-Thiouridine, is a modified nucleotide of tRNA that is conserved from bacteria to archaea, where it functions as a sensor for near-UV irradiation tRNA-4-thiouridine
-
 ?

Organism

Organism UniProt Comment Textmining
Methanococcus maripaludis Q6LXJ6
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Methanococcus maripaludis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
sulfide + tRNA-uridine the enzyme is invoved in biosynthesis of 4-thiouridine in tRNA. 4-Thiouridine, is a modified nucleotide of tRNA that is conserved from bacteria to archaea, where it functions as a sensor for near-UV irradiation Methanococcus maripaludis tRNA-4-thiouridine
-
 ?
sulfide + [tRNACys]-uridine proposed mechanism: first, a sulfur donor (equivalent of S2-) attacks the Cys265–Cys268 disulfide linkage to generate a persulfide on either Cys265 or Cys268, leaving the other Cys as a free thiol. Then, a thiolate derived from deprotonation of the free thiol attacks the bridging sulfur of the persulfide to liberate the terminal sulfur. After donation of the sulfur (in the -2 oxidation state) to form 4-thiouridine, the disulfide (with both sulfurs in the -1 oxidation state) is consequently regenerated. No exogenous electron donor or acceptor is required for the catalytic cycle Methanococcus maripaludis [tRNACys]-4-thiouridine
-
 ?

Synonyms

Synonyms Comment Organism
MMP1354 locus name Methanococcus maripaludis
ThiI
-
 Methanococcus maripaludis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Methanococcus maripaludis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.3
-
 assay at Methanococcus maripaludis

Cofactor

Cofactor Comment Organism Structure
additional information the enzyme requires two conserved Cys residues (Cys265 and Cys268), which are arranged in a CXXC motif in the PP-loop domain, for both in vitro and in vivo activities to generate 4-thiouridine in tRNAs Methanococcus maripaludis

General Information

General Information Comment Organism
malfunction deletion mutant shows no groth defects compared to wild-type, indicating that MMP1354 is not an essential enzyme Methanococcus maripaludis
malfunction deletion of ThiI abolishes the biosynthesis of 4-thiouridine but not of thiamine Methanococcus maripaludis
malfunction MMP1354 complements an Escherichia coli Thi mutant Methanococcus maripaludis