Crystallization (Comment) | Organism |
---|---|
purified isozyme MPH(2')-I in apoform and in complex with GTP analogues and six different macrolides (guanosine 5'-[beta,gamma-imido]triphosphate (GMPPNP) or guanosine diphosphate (GDP) and either a 14-membered lactone macrolide: erythromycin, oleandomycin, or clarithromycin, or the 15-membered lactone macrolide azithromycin), X-ray diffraction structure determination analysis at 1.2-1.6 A resolution, molecular replacement method, modeling. Despite the inclusion of MgCl2 in the crystallization experiments, no evidence of magnesium ions is observed in any of the MPH(2')-I electron density maps | Escherichia coli |
purified isozyme MPH(2')-II in apoform and in complex with GTP analogues and six different macrolides (guanosine diphosphate (GDP) or GDP or GTPgS (guanosine 5'-O-[gamma-thio]triphosphate) and either a 14-membered lactone macrolide: erythromycin, oleandomycin, or clarithromycin, or the 15-membered lactone macrolide azithromycin), X-ray diffraction structure determination analysis at 1.31-1.65 A resolution, molecular replacement method, modeling. At least one divalent metal ion (either Mg2+ or Ca2+, based on the geometry and bond distances) from the crystallization solution is observed in the nucleotide binding pocket | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + oleandomycin | Escherichia coli | - |
ADP + oleandomycin 2'-O-phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | O32553 | - |
- |
Escherichia coli | Q9EVJ6 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + oleandomycin | - |
Escherichia coli | ADP + oleandomycin 2'-O-phosphate | - |
? | |
additional information | structure-based substrate specificity, macrolide binding analysis, overview. Although erythromycin, clarithromycin, oleandomycin, and azithromycin differ in the size of the lactone ring and varying substitutions, the 14- and 15-membered ring substrates bind to the two MPH enzymes in a similar location, adopt similar conformations, and interact with the enzymes in a similar manner. MPH(2')-I cannot modify 16-membered macrolides, in contrast to MPH(2')-II | Escherichia coli | ? | - |
- |
|
additional information | structure-based substrate specificity, macrolide binding analysis, overview. Although erythromycin, clarithromycin, oleandomycin, and azithromycin differ in the size of the lactone ring and varying substitutions, the 14- and 15-membered ring substrates bind to the two MPH enzymes in a similar location, adopt similar conformations, and interact with the enzymes in a similar manner. Unlike MPH(2')-I, MPH(2')-II can efficiently modify 16-membered macrolides. The 16-membered rings are more ovoid in shape compared with the rounded shape of the 14- and 15-membered macrolides. The interactions between MPH(2')-II and the 16-membered macrolides, spiramycin and josamycin, are similar to those of the 14- and 15-membered macrolides, involving many of the same amino acid side chains | Escherichia coli | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
macrolide 2'-phosphotransferase type I | - |
Escherichia coli |
macrolide 2'-phosphotransferase type II | - |
Escherichia coli |
MPH(2')-I | - |
Escherichia coli |
MPH(2')-II | - |
Escherichia coli |
mphA | - |
Escherichia coli |
mphB | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
evolution | the macrolide phosphotransferase structures show that the enzymes are related to the aminoglycoside phosphotransferases, but are distinguished from them by the presence of a large interdomain linker that contributes to an expanded antibiotic binding pocket | Escherichia coli |
additional information | the large interdomain linker contributes to an expanded antibiotic binding pocket. This pocket is largely hydrophobic, with a negatively charged patch located at a conserved aspartate residue. Comparison of the enzyme-macrolide complex structure with the structures of macrolides bound to their natural target, the 50S ribosome, overview. Nucleotides bind to MPH(2')-I in a cleft between the N-terminal lobe and the core subdomain of the C-terminal lobe, binding site structure analysis | Escherichia coli |
additional information | the large interdomain linker contributes to an expanded antibiotic binding pocket. This pocket is largely hydrophobic, with a negatively charged patch located at a conserved aspartate residue. Comparison of the enzyme-macrolide complex structure with the structures of macrolides bound to their natural target, the 50S ribosome, overview. Nucleotides bind to MPH(2')-II in a cleft between the N-terminal lobe and the core subdomain of the C-terminal lobe, binding site structure analysis | Escherichia coli |
physiological function | macrolide phosphotransferase enzymes can inactivate the macrolides, a class of antibiotic, characterized by a large macrocyclic lactone ring. Broad-spectrum resistance is conferred by the enzymes | Escherichia coli |