Application | Comment | Organism |
---|---|---|
drug development | the enzyme can aid in the development of cell wall targeting therapeutics against Mycobacterium tuberculosis | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
gene Rv3717, expression of recombinant GST-tagged enzyme in Escherichia coli strain BL-21 Codon Plus | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
purified enzyme Rv3717 free, or in complex with L-Ala-iso-D-Gln product or Zn2+, hanging drop vapor diffusion method, from 200 mM NaCl, 100 mM Tris, pH 8.5, 25% PEG 3350, X-ray diffraction structure determination and analysis at 2.1-2.67 A resolution, molecular replacement method using Bartonella henselae AmiB as the search model, PDB code 3NE8 | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
E200X | inactive mutant | Mycobacterium tuberculosis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | Zn2 is required for hydrolysis of muramyl dipeptide, Zn2 binding rearranges active site residues. The catalytic residue Glu200 does not participate in binding to Zn2+ ions | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | I6Y4D2 | - |
- |
Mycobacterium tuberculosis H37Rv | I6Y4D2 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged enzyme from Escherichia coli strain BL-21 Codon Plus by glutathione affinity chromatography and tag cleavage, followed by a combined glutathione affinity/anion exchange chromatography step, and a gel filtration step | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | no activity on polymerized peptidoglycan | Mycobacterium tuberculosis | ? | - |
? | |
additional information | no activity on polymerized peptidoglycan | Mycobacterium tuberculosis H37Rv | ? | - |
? | |
MurNAc-L-Ala-D-isoGln + H2O | - |
Mycobacterium tuberculosis | N-acetylmuramic acid + L-alanyl-iso-D-glutamine | - |
? | |
MurNAc-L-Ala-D-isoGln + H2O | - |
Mycobacterium tuberculosis H37Rv | N-acetylmuramic acid + L-alanyl-iso-D-glutamine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Mtb peptidoglycan amidase | - |
Mycobacterium tuberculosis |
Rv3717 | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the Pfam family Amidase_3, the mode of substrate binding is likely shared by the Amidase_3 family proteins | Mycobacterium tuberculosis |
additional information | the enzyme contains an extra disulfide-bonded beta-hairpin adjacent to the active site compared to other peptidoglycan amidases. Zn2 binding rearranges active site residues and disulfide formation promotes folding of the beta-hairpin. Although Zn2 is required for hydrolysis of muramyl dipeptide, disulfide oxidation is not required for activity on this substrate. The product L-alanine-iso-D-glutamine binds at the head of a closed tunnel. The orientation of the product in the active site suggests a role for a conserved Glu200 in catalysis | Mycobacterium tuberculosis |
physiological function | potential role for enzyme Rv3717 in peptidoglycan fragment recycling | Mycobacterium tuberculosis |