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Literature summary extracted from

  • Dawson, C.D.; Irwin, S.M.; Backman, L.R.F.; Le, C.; Wang, J.X.; Vennelakanti, V.; Yang, Z.; Kulik, H.J.; Drennan, C.L.; Balskus, E.P.
    Molecular basis of C-S bond cleavage in the glycyl radical enzyme isethionate sulfite-lyase (2021), Cell Chem. Biol., 28, 1-14 .
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.4.1.38 structure in complex with isethionate, to 2.7 A resolution. In comparison with other glycyl radical enzymes, alternate positioning of the active site beta strands allows for distinct residue positions to contribute to substrate binding, thus creating a highly tailored active site for the binding of the negatively charged isethionate substrate Bilophila wadsworthia

Protein Variants

EC Number Protein Variants Comment Organism
4.4.1.38 C468S variant can form a glycyl radical but has no detectable sulfite release in an endpoint assay Bilophila wadsworthia
4.4.1.38 E470Q variant can partly be activated by IslB, displays no endpoint activity Bilophila wadsworthia
4.4.1.38 F682A decrease in catalytic efficiency by 1-2 orders of magnitude Bilophila wadsworthia
4.4.1.38 F682Y decrease in catalytic efficiency by 1-2 orders of magnitude Bilophila wadsworthia
4.4.1.38 G805A residue Gly805 forms the glycyl radical Bilophila wadsworthia
4.4.1.38 I192A mutant has nearly identical radical installation to the wild-type, and a similar KM with a decreased kcat value Bilophila wadsworthia
4.4.1.38 Q193A variant can partly be activated by IslB, displays no endpoint activity Bilophila wadsworthia
4.4.1.38 R189E variant can partly be activated by IslB, displays no endpoint activity Bilophila wadsworthia
4.4.1.38 R189E/R678E variant can partly be activated by IslB, displays no endpoint activity Bilophila wadsworthia
4.4.1.38 R678E variant can partly be activated by IslB, displays no endpoint activity Bilophila wadsworthia
4.4.1.38 V680A mutant has the maximal amount of glycyl radical installation of the assayed mutants despite being inactive toward isethinate Bilophila wadsworthia
4.4.1.38 W374F decrease in catalytic efficiency by 1-2 orders of magnitude Bilophila wadsworthia
4.4.1.38 W374Y decrease in catalytic efficiency by 1-2 orders of magnitude Bilophila wadsworthia

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.4.1.38 6.8
-
 isethionate mutant F682A, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 8
-
 isethionate wild-type, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 8
-
 isethionate mutant F682Y, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 11
-
 isethionate mutant W374F, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 12
-
 isethionate mutant I192A, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 16
-
 isethionate mutant W374Y, pH 7.5, 30°C Bilophila wadsworthia

Organism

EC Number Organism UniProt Comment Textmining
4.4.1.38 Bilophila wadsworthia E5Y378
-
-
4.4.1.38 Bilophila wadsworthia 3_1_6 E5Y378
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.4.1.38 isethionate
-
 Bilophila wadsworthia acetaldehyde + sulfite
-
 ?
4.4.1.38 isethionate
-
 Bilophila wadsworthia 3_1_6 acetaldehyde + sulfite
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.4.1.38 0.023
-
 isethionate mutant F682Y, pH 7.5, 30°C, glycyl radical normalized value Bilophila wadsworthia
4.4.1.38 0.133
-
 isethionate mutant W374F, pH 7.5, 30°C, glycyl radical normalized value Bilophila wadsworthia
4.4.1.38 0.154
-
 isethionate mutant F682A, pH 7.5, 30°C, glycyl radical normalized value Bilophila wadsworthia
4.4.1.38 0.465
-
 isethionate mutant W374Y, pH 7.5, 30°C, glycyl radical normalized value Bilophila wadsworthia
4.4.1.38 1.9
-
 isethionate mutant I192A, pH 7.5, 30°C, glycyl radical normalized value Bilophila wadsworthia
4.4.1.38 9.5
-
 isethionate wild-type, pH 7.5, 30°C, glycyl radical normalized value Bilophila wadsworthia

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.4.1.38 0.0029
-
 isethionate mutant F682Y, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 0.012
-
 isethionate mutant W374F, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 0.023
-
 isethionate mutant F682A, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 0.029
-
 isethionate mutant W374Y, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 0.16
-
 isethionate mutant I192A, pH 7.5, 30°C Bilophila wadsworthia
4.4.1.38 1.2
-
 isethionate wild-type, pH 7.5, 30°C Bilophila wadsworthia