Application | Comment | Organism |
---|---|---|
drug development | the human pathogen Mycobacterium tuberculosis PPTases are attractive drug targets | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
gene Rv2794c or pptT, recombinant expression of MBP-tagged type II PPTase PptT in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant MBP-tagged type-II PPTase PptT, hanging drop vapor diffusion, mixing of 0.002 ml of 10 mg/ml protein in 20 mM MES, pH 6.7, 300 mM NaCl, 10% v/v glycerol, 0.5 mM tris(2-carboxyethyl)phosphine, and 1 mM CoA, with 0.001 ml of reservoir solution containing 1.6 M sodium citrate, 18°C, method optimization, X-ray diffraction structure determination and analysis at 1.75 A resolution, modeling. No crystals can be grown without added CoA. Recombinantly expressed enzyme PptT as N- or C-terminally His6-tagged proteins are soluble protein in Escherichia coli, but both are unstable over time and neither can be crystallized | Mycobacterium tuberculosis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | dependent on, binding structure, overview. The edge strands beta4 and beta6 of the two beta-sheets provide the binding site for the CoA diphosphate and associated Mg2+ ion. The active site in Mtb-PptT is formed in the shallow cleft between the two domains, where CoA and Mg2+ are bound | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
CoA-[4'-phosphopantetheine] + apo-[acyl-carrier protein] | Mycobacterium tuberculosis | - |
adenosine 3',5'-bisphosphate + holo-[acyl-carrier protein] | - |
? | |
CoA-[4'-phosphopantetheine] + apo-[acyl-carrier protein] | Mycobacterium tuberculosis ATCC 25618 | - |
adenosine 3',5'-bisphosphate + holo-[acyl-carrier protein] | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | O33336 | - |
- |
Mycobacterium tuberculosis ATCC 25618 | O33336 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant MBP-tagged type II PPTase PptT from Escherichia coli strain BL21(DE3) amylose affinity chromatography, anion exchange chromatography, and gel filtration | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
CoA-[4'-phosphopantetheine] + apo-[acyl-carrier protein] | - |
Mycobacterium tuberculosis | adenosine 3',5'-bisphosphate + holo-[acyl-carrier protein] | - |
? | |
CoA-[4'-phosphopantetheine] + apo-[acyl-carrier protein] | The edge strands beta4 and beta6 of the two beta-sheets provide the binding site for the CoA diphosphate and associated Mg2+ ion. The active site in Mtb-PptT is formed in the shallow cleft between the two domains, where CoA and Mg2+ are bound | Mycobacterium tuberculosis | adenosine 3',5'-bisphosphate + holo-[acyl-carrier protein] | - |
? | |
CoA-[4'-phosphopantetheine] + apo-[acyl-carrier protein] | - |
Mycobacterium tuberculosis ATCC 25618 | adenosine 3',5'-bisphosphate + holo-[acyl-carrier protein] | - |
? | |
CoA-[4'-phosphopantetheine] + apo-[acyl-carrier protein] | The edge strands beta4 and beta6 of the two beta-sheets provide the binding site for the CoA diphosphate and associated Mg2+ ion. The active site in Mtb-PptT is formed in the shallow cleft between the two domains, where CoA and Mg2+ are bound | Mycobacterium tuberculosis ATCC 25618 | adenosine 3',5'-bisphosphate + holo-[acyl-carrier protein] | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | Mycobacterium tuberculosis type-II PPTase PptT reveals an alpha/beta fold broadly similar to other type-II PPTases, but with differences in peripheral structural elements. Three-dimensional structure of Mtb-PptT, modeling, overview. Mtb-PptT comprises two alpha/beta domains with pseudo 2fold symmetry | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
phosphopantetheinyl transferase | - |
Mycobacterium tuberculosis |
PptT | - |
Mycobacterium tuberculosis |
PTase | - |
Mycobacterium tuberculosis |
Rv2794c | - |
Mycobacterium tuberculosis |
type-II PPTase PptT | - |
Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
additional information | the human pathogen Mycobacterium tuberculosis encodes two PPTases, a type-I PPTase and a type-II PPTase, that are both essential. Type-II PPTase PptT shows a bound CoA that is clearly defined with its pantetheinyl arm tucked into a hydrophobic pocket. Interactions involving the CoA diphosphate, bound Mg2+ and three active site acidic side chains suggest a plausible pathway for proton transfer during catalysis. Three-dimensional structure of Mtb-PptT, modeling, overview. Mtb-PptT comprises two alpha/beta domains with pseudo 2fold symmetry. It is octahedrally coordinated to the alpha- and beta-phosphates, Asp114, Glu116, the peptide oxygen of Ala115 and a water molecule | Mycobacterium tuberculosis |