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Literature summary extracted from

  • Gengenbacher, M.; Xu, T.; Niyomrattanakit, P.; Spraggon, G.; Dick, T.
    Biochemical and structural characterization of the putative dihydropteroate synthase ortholog Rv1207 of Mycobacterium tuberculosis (2008), FEMS Microbiol. Lett., 287, 128-135.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.5.1.15 drug development cannot bypass DHPS activity during inhibition of the functional ortholog MtDHPS (folP1) as treatment of tuberculosis Mycobacterium tuberculosis
2.5.1.15 drug development target in treatment of tuberculosis Mycobacterium tuberculosis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.15 from genomic DNA (strain H37Rv) in pET-21a for expression with C-terminal hexa-His tag in Escherichia coli BL21 (DE3) Mycobacterium tuberculosis
2.5.1.15 from genomic DNA (strain H37Rv) in pQE60 for expression with C-terminal hexa-His tag in Escherichia coli M15 Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.15 PDB: 2VP8, conforms classical triosephosphate isomerase (TIM) barrel arrangement, 6-hydroxymethyl-7,8-dihydropterin-diphosphate-binding pocket occupied by the histidine 220 side chain compared to leucine 180 in the functional ortholog MtDHPS (PDB: 1EYE), crystals: space group P3(1)21, 2 molecules per asymmetric unit, noncrystallographic dimer in the crystal structure, unit cell parameters: a, b: 80.73, c: 215.94, hanging drop vapour method: 25% ethylene glycole + protein solution (16 mg/ml), 2 days, 18°C Mycobacterium tuberculosis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.15 29000
-
 recombinant, hexa-His tagged enzyme, SDS-PAGE Mycobacterium tuberculosis
2.5.1.15 33000
-
 recombinant, hexa-His tagged enzyme, SDS-PAGE Mycobacterium tuberculosis
2.5.1.15 34070
-
 deduced from primary structure Mycobacterium tuberculosis
2.5.1.15 75000
-
 recombinant hexa-His tagged enzyme, gel filtration chromatography Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.15 Mycobacterium tuberculosis P9WNC9
-
-
2.5.1.15 Mycobacterium tuberculosis P9WND1
-
-
2.5.1.15 Mycobacterium tuberculosis H37Rv P9WNC9
-
-
2.5.1.15 Mycobacterium tuberculosis H37Rv P9WND1
-
-
2.5.1.15 Mycobacterium tuberculosis variant bovis A0A0H3MCB3
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.15 recombinant, hexa-His tagged enzyme from bacterial lysate by nickel-nitrilotriacetic acid agarose, elution with 200 mM imidazole, stored at -80°C in 10% glycerol or further purified on by Superdex 200 16/60 gel filtration chromatography Mycobacterium tuberculosis
2.5.1.15 recombinant, hexa-His tagged enzyme from bacterial lysate by nickel-nitrilotriacetic acid agarose, elution with 200 mM imidazole, stored at -80°C in 10% glycerol or further purified on by Superdex 200 GL 10/300 gel filtration chromatography Mycobacterium tuberculosis

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.5.1.15 cell suspension culture logarithmic growing bacteria, Western blotting Mycobacterium tuberculosis variant bovis
-
2.5.1.15 cell suspension culture qiescent bacteria, lower expression level under anaerobic conditions compared to nutrient starvation, Western blotting Mycobacterium tuberculosis variant bovis
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.15 0.005
-
 +/-3, assumed inactive because 100times less than folP1 gene product Mycobacterium tuberculosis
2.5.1.15 0.577
-
 +/-12, pH 9 Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.15 additional information binding of 6-hydroxymethyl-7,8-dihydropterin-diphosphate with KD: 117 microM, revealed by fluorescence spectroscopy Mycobacterium tuberculosis ?
-
 ?
2.5.1.15 additional information binding of 6-hydroxymethyl-7,8-dihydropterin-diphosphate with KD: 4.6 microM, revealed by fluorescence spectroscopy Mycobacterium tuberculosis ?
-
 ?
2.5.1.15 additional information binding of dapsone (p-aminobenzoate substrate analog) with KD: 168 microM, in absence of 6-hydroxymethyl-7,8-dihydropterin-diphosphate, , revealed by fluorescence spectroscopy Mycobacterium tuberculosis ?
-
 ?
2.5.1.15 additional information binding of dapsone (p-aminobenzoate substrate analog) with KD: 96 microM, in absence of 6-hydroxymethyl-7,8-dihydropterin-diphosphate, revealed by fluorescence spectroscopy Mycobacterium tuberculosis ?
-
 ?
2.5.1.15 additional information Rv1207 lacks dihydropteroate synthase activity in presence of 10 microM p-aminobenzoate and 10 microM 6-hydroxymethyl-7,8-dihydropterin-diphosphate, 5 mM MgCl2, 1 mM dithiothreitol, 37°C, pH 7-9.5 Mycobacterium tuberculosis ?
-
 ?
2.5.1.15 additional information binding of 6-hydroxymethyl-7,8-dihydropterin-diphosphate with KD: 117 microM, revealed by fluorescence spectroscopy Mycobacterium tuberculosis H37Rv ?
-
 ?
2.5.1.15 additional information binding of dapsone (p-aminobenzoate substrate analog) with KD: 96 microM, in absence of 6-hydroxymethyl-7,8-dihydropterin-diphosphate, revealed by fluorescence spectroscopy Mycobacterium tuberculosis H37Rv ?
-
 ?
2.5.1.15 additional information Rv1207 lacks dihydropteroate synthase activity in presence of 10 microM p-aminobenzoate and 10 microM 6-hydroxymethyl-7,8-dihydropterin-diphosphate, 5 mM MgCl2, 1 mM dithiothreitol, 37°C, pH 7-9.5 Mycobacterium tuberculosis H37Rv ?
-
 ?
2.5.1.15 additional information binding of 6-hydroxymethyl-7,8-dihydropterin-diphosphate with KD: 4.6 microM, revealed by fluorescence spectroscopy Mycobacterium tuberculosis H37Rv ?
-
 ?
2.5.1.15 additional information binding of dapsone (p-aminobenzoate substrate analog) with KD: 168 microM, in absence of 6-hydroxymethyl-7,8-dihydropterin-diphosphate, , revealed by fluorescence spectroscopy Mycobacterium tuberculosis H37Rv ?
-
 ?
2.5.1.15 p-aminobenzoate + 6-hydroxymethyl-7,8-dihydropterin-diphosphate 5 mM MgCl2, 1 mM dithiothreitol, 37°C, pH 7-9.5 Mycobacterium tuberculosis diphosphate + 7,8-dihydropteroate
-
 ?
2.5.1.15 p-aminobenzoate + 6-hydroxymethyl-7,8-dihydropterin-diphosphate 5 mM MgCl2, 1 mM dithiothreitol, 37°C, pH 7-9.5 Mycobacterium tuberculosis H37Rv diphosphate + 7,8-dihydropteroate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.5.1.15 homodimer 2 * 34070, gel filtration chromatography Mycobacterium tuberculosis

Synonyms

EC Number Synonyms Comment Organism
2.5.1.15 BCG3672c
-
 Mycobacterium tuberculosis variant bovis
2.5.1.15 DHPS
-
 Mycobacterium tuberculosis
2.5.1.15 folP1
-
 Mycobacterium tuberculosis
2.5.1.15 folP2
-
 Mycobacterium tuberculosis
2.5.1.15 MbDHPS
-
 Mycobacterium tuberculosis variant bovis
2.5.1.15 MtDHPS
-
 Mycobacterium tuberculosis
2.5.1.15 Mycobacterium tuberculosis dihydropteroate synthase
-
 Mycobacterium tuberculosis
2.5.1.15 putative dihydropteroate synthase ortholog
-
 Mycobacterium tuberculosis
2.5.1.15 Rv1207
-
 Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.15 9
-
 according to specific activity Mycobacterium tuberculosis