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Literature summary extracted from

  • Wheatley, R.W.; Zheng, R.B.; Richards, M.R.; Lowary, T.L.; Ng, K.K.
    Tetrameric structure of the GlfT2 galactofuranosyltransferase reveals a scaffold for the assembly of mycobacterial arabinogalactan (2012), J. Biol. Chem., 287, 28132-28143.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.288 expressed in Escherichia coli C41(DE3) cells Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.288 hanging drop vapor diffusion method, free enzyme is crystallized using 5-7% (w/v) PEG 4000, 100 mM sodium cacodylate, pH 6.5, 100 mM sodium acetate. UDP-bound enzyme is crystallized using 6-7% (w/v) PEG 4000, 100 mM sodium acetate, 10% (v/v) glycerol, 5 mM TCEP, at 22°C Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.288 D372S the mutant shows no detectable activity Mycobacterium tuberculosis
2.4.1.288 E300S the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme Mycobacterium tuberculosis
2.4.1.288 E371S the mutant shows no detectable activity Mycobacterium tuberculosis
2.4.1.288 H413S the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme Mycobacterium tuberculosis
2.4.1.288 W399S the mutant exhibits increased Km and strongly decreased kcat values compare to the wild type enzyme Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.288 0.38
-
 UDP-alpha-D-galactofuranose wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.41
-
 UDP-alpha-D-galactofuranose mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.42
-
 UDP-alpha-D-galactofuranose mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.43
-
 UDP-alpha-D-galactofuranose mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.6
-
 beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3 wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 1.54
-
 beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3 mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 1.78
-
 beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3 mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 1.78
-
 beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3 mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.288 Mn2+ Mn2+ is bound to the active site Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.288 Mycobacterium tuberculosis O53585
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.288 nickel-Sepharose column chromatography and Sephacryl S-200 gel filtration Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.288 UDP-alpha-D-galactofuranose + beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->4)-alpha-L-rhamnopyranosyl-(1->3)-N-acetyl-alpha-D-glucosaminyl-diphospho-trans,octacis-decaprenol the beta-(1->5)-transferase activity of the enzyme first glycosylates the galactofuranosyl residue at the nonreducing terminus of the polyprenol-bound tetrasaccharide acceptor. The resulting product is then a substrate for the beta-(1->6)-transferase activity of the enzyme Mycobacterium tuberculosis UDP + [beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->6)]14-beta-D-galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->4)-alpha-L-rhamnopyranosyl-(1->3)-N-acetyl-alpha-D-glucosaminyl-diphospho-trans,octacis-decaprenol
-
 ?
2.4.1.288 UDP-alpha-D-galactofuranose + beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
-
 Mycobacterium tuberculosis UDP + ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.1.288 homotetramer x-ray crystallography Mycobacterium tuberculosis

Synonyms

EC Number Synonyms Comment Organism
2.4.1.288 galactofuranosyltransferase
-
 Mycobacterium tuberculosis
2.4.1.288 GlfT2
-
 Mycobacterium tuberculosis
2.4.1.288 Rv3808c
-
 Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.288 0.004
-
 UDP-alpha-D-galactofuranose mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.004
-
 beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3 mutant enzyme H413S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.005
-
 UDP-alpha-D-galactofuranose mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.005
-
 beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3 mutant enzyme E300S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.006
-
 UDP-alpha-D-galactofuranose mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 0.006
-
 beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3 mutant enzyme W399S, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 7.2
-
 UDP-alpha-D-galactofuranose wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis
2.4.1.288 7.2
-
 beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3 wild type enzyme, in 50 mM MOPS, pH 7.6, 50 mM KCl, 20 mM MgCl2, at 37°C Mycobacterium tuberculosis

General Information

EC Number General Information Comment Organism
2.4.1.288 physiological function GlfT2 is essential for mycobacterial cell wall biosynthesis Mycobacterium tuberculosis