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Literature summary for 2.4.1.288 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
Search for more literature for 2.4.1.288

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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
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
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
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
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
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

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

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis O53585
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
UDP-alpha-D-galactofuranose + beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-O(CH2)7CH3
-
 Mycobacterium tuberculosis UDP + ?
-
 ?

Subunits

Subunits Comment Organism
homotetramer x-ray crystallography Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
galactofuranosyltransferase
-
 Mycobacterium tuberculosis
GlfT2
-
 Mycobacterium tuberculosis
Rv3808c
-
 Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
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
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
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
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
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
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
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

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