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Literature summary for 2.7.7.71 extracted from

  • Kim, S.; Kim, M.S.; Jo, S.; Shin, D.H.
    GTP preference of D-glycero-alpha-D-manno-heptose-1-phosphate guanylyltransferase from Yersinia pseudotuberculosis (2019), Int. J. Mol. Sci., 21, 280 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.71

Cloned(Commentary)

Cloned (Comment) Organism
gene hddC, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Yersinia pseudotuberculosis

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant wild-type His6-tagged enzyme with bound GMPPNP and Mg2+, hanging drop vapor diffusion method, from 0.05 M HEPES, pH 7.5, 1.2 M sodium citrate tribasic dehydrate, and 0.2 M MgCl2, room temperature, method optimization, X-ray diffraction structure determination and analysis at 1.55 A resolution, molecular replacement using the coordinates of native YpHddC (PDB ID 5XHW) as a search model Yersinia pseudotuberculosis

Protein Variants

Protein Variants Comment Organism
S53N site-directed mutagenesis, the mutant is constructed to mimic the UTP binding site of PpMurU, but GTP is still the major cofactor for the nucleotidylyltransferase activity for the mutant Yersinia pseudotuberculosis
S53V/E80Q site-directed mutagenesis, the mutant is constructed to mimic the nucleotide-binding pocket of the UDPGP family, the GTP affinity is severely diminished without increment of pyrimidine affinity Yersinia pseudotuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ magnesium is absolutely required for catalytic activity of YpHddC as a cofactor and also is essential for crystallization Yersinia pseudotuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-glycero-alpha-D-manno-heptose 1-phosphate + GTP Yersinia pseudotuberculosis
-
 GDP-D-glycero-alpha-D-manno-heptose + diphosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Yersinia pseudotuberculosis Q8GJ90 serotype I
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Yersinia pseudotuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-glycero-alpha-D-manno-heptose 1-phosphate + GTP
-
 Yersinia pseudotuberculosis GDP-D-glycero-alpha-D-manno-heptose + diphosphate
-
 ?

Synonyms

Synonyms Comment Organism
D-glycero-alpha-D-manno-heptose-1-phosphate guanylyltransferase
-
 Yersinia pseudotuberculosis
HddC
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 Yersinia pseudotuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Yersinia pseudotuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Yersinia pseudotuberculosis

General Information

General Information Comment Organism
metabolism D-glycero-alpha-D-manno-heptose-1-phosphate guanylyltransferase (HddC) is the fourth enzyme synthesizing a building component of lipopolysaccharide (LPS) of Gram-negative bacteria Yersinia pseudotuberculosis
additional information manual docking of GTP with a native YpHddC. Glu80 is found in a unique motif EXEPLGTGGA among approximately five thousand HddC homologues. Therefore, the formation of hydrogen bonds between two oxygens of Glu80 and two nitrogen atoms of GMPPN is suggests to be a major driving force to select and interact with the guanine ring of GMPPN. Glu80 is the key residue to hold GTP in the nucleotide-binding pocket. The active site residues of YpHddC are properly designed to accommodate GTP as a crucial base Yersinia pseudotuberculosis