Characterization of the active site and insight into the binding mode of the anti-angiogenesis agent fumagillin to the manganese(II)-loaded methionyl aminopeptidase from Escherichia coli

@article{Dsouza2004CharacterizationOT,
  title={Characterization of the active site and insight into the binding mode of the anti-angiogenesis agent fumagillin to the manganese(II)-loaded methionyl aminopeptidase from Escherichia coli},
  author={V. D'souza and R. Brown and B. Bennett and R. Holz},
  journal={JBIC Journal of Biological Inorganic Chemistry},
  year={2004},
  volume={10},
  pages={41-50}
}
EPR spectra were recorded for methionine aminopeptidase from Escherichia coli (EcMetAP-I) samples (~2.5 mM) to which one and two equivalents of Mn(II) were added (the latter is referred to as [MnMn(EcMetAP-I)]). The spectra for each sample were indistinguishable except that the spectrum of [MnMn(EcMetAP-I)] was twice as intense. The EPR spectrum of [MnMn(EcMetAP-I)] exhibited the characteristic six-line g≈2 EPR signal of mononuclear Mn(II) with Aav(55Mn)=9.3 mT (93 G) and exhibited Curie-law… Expand
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References

SHOWING 1-10 OF 49 REFERENCES
Spectroscopically distinct cobalt(II) sites in heterodimetallic forms of the aminopeptidase from Aeromonas proteolytica: characterization of substrate binding.
TLDR
It is established that substrate binds only to the first metal binding site in AAP and thus substantiates the first step in catalysis in the recently proposed mechanism of action for AAP. Expand
Kinetic and structural characterization of manganese(II)-loaded methionyl aminopeptidases.
TLDR
Manganese(II) activation of the methionyl aminopeptidases from Escherichia coli and the hyperthermophilic archaeon Pyrococcus furiosus (PfMetAP-II) was investigated and specific activity and K(m) values increased with increasing temperature. Expand
Spectroscopic identification of a dinuclear metal centre in manganese(II)-activated aminopeptidase P from Escherichia coli: implications for human prolidase
Abstract Electron paramagnetic resonance (EPR) spectra and X-ray absorption (EXAFS and XANES) data have been recorded for the manganese enzyme aminopeptidase P (AMPP, PepP protein) from EscherichiaExpand
The anti-angiogenic agent fumagillin covalently modifies a conserved active-site histidine in the Escherichia coli methionine aminopeptidase.
TLDR
Fumagillin and ovalicin covalently modify a conserved histidine residue in the active site of the MetAP from Escherichia coli, a type I enzyme, suggesting that these compounds may be effective pharmacological agents against pathogenic and resistant forms of E. coli and other microorganisms. Expand
Structural evidence that the methionyl aminopeptidase from Escherichia coli is a mononuclear metalloprotease.
TLDR
The EXAFS data suggest that a dinuclear Co(II) center does not exist in EcMetAP and that the first Co atom is located in the histidine-ligated side of the active site. Expand
Divalent metal binding properties of the methionyl aminopeptidase from Escherichia coli.
TLDR
The metal-binding properties of the methionyl aminopeptidase from Escherichia coli (MetAP) were investigated and measurements of catalytic activity revealed that maximal enzymatic activity is observed after the addition of only 1 equiv of divalent metal ion. Expand
The 1.15A crystal structure of the Staphylococcus aureus methionyl-aminopeptidase and complexes with triazole based inhibitors.
TLDR
The 1.15A crystal structure of the Staphylococcus aureus MetAP-I as an apo-enzyme and its complexes with various 1,2,4-triazole-based derivatives at high-resolution is described. Expand
L-arginine binding to liver arginase requires proton transfer to gateway residue His141 and coordination of the guanidinium group to the dimanganese(II,II) center.
TLDR
The present EPR studies reveal that there is no essential alteration of the Mn2 site upon mutation of His141 to an Asn residue, which lacks a potential acid/base residue, while the catalytic activity of the mutant enzyme is 10 times lower vs wild-type enzyme. Expand
Physiologically relevant metal cofactor for methionine aminopeptidase-2 is manganese.
TLDR
It is concluded that MetAP2 is a manganese enzyme and that therapeutic Met AP2 inhibitors should inhibit MetAP1-Mn(2+), and which metal ion is physiologically relevant. Expand
Insights into the mechanism of Escherichia coli methionine aminopeptidase from the structural analysis of reaction products and phosphorus-based transition-state analogues.
TLDR
Observations tend to support one of the mechanistic schemes for eMetAP considered before, although with a revision in the role played by His79, and suggest parallels between the mechanism of action of methionine aminopeptidase and other "pita-bread" enzymes. Expand
...
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4
5
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