Molecular recognition of angiogenesis inhibitors fumagillin and ovalicin by methionine aminopeptidase 2.
Angiogenesis inhibitors are a novel class of promising therapeutic agents for treating cancer and other human diseases. Fumagillin and ovalicin compose a class of structurally related natural products that potently inhibit angiogenesis by blocking endothelial cell proliferation. A synthetic analog of fumagillin, TNP-470, is currently undergoing clinical trials for treatment of a variety of cancers. A common target for fumagillin and ovalicin recently was identified as the type 2 methionine aminopeptidase (MetAP2). These natural products bind MetAP2 covalently, inhibiting its enzymatic activity. The specificity of this binding is underscored by the lack of inhibition of the closely related type 1 enzyme, MetAP1. The molecular basis of the high affinity and specificity of these inhibitors for MetAP2 has remained undiscovered. To determine the structural elements of these inhibitors and MetAP2 that are involved in this interaction, we synthesized fumagillin analogs in which each of the potentially reactive epoxide groups was removed either individually or in combination. We found that the ring epoxide in fumagillin is involved in the covalent modification of MetAP2, whereas the side chain epoxide group is dispensable. By using a fumagillin analog tagged with fluorescein, His-231 in MetAP2 was identified as the residue that is covalently modified by fumagillin. Site-directed mutagenesis of His-231 demonstrated its importance for the catalytic activity of MetAP2 and confirmed that the same residue is covalently modified by fumagillin. These results, in agreement with a recent structural study, suggest that fumagillin and ovalicin inhibit MetAP2 by irreversible blockage of the active site.
Griffith
E C
EC
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Su
Z
Z
Niwayama
S
S
Ramsay
C A
CA
Chang
Y H
YH
Liu
J O
JO
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology; D020134:Catalytic Domain; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003510:Cyclohexanes; D003545:Cysteine; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D005231:Fatty Acids, Unsaturated / Q000737:chemistry / Q000494:pharmacology*; D006639:Histidine; D006801:Humans; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D012717:Sesquiterpenes / Q000737:chemistry / Q000494:pharmacology*; D018411:Spodoptera; D014162:Transfection
D000900:Anti-Bacterial Agents; D003510:Cyclohexanes; D005231:Fatty Acids, Unsaturated; D011994:Recombinant Proteins; D012717:Sesquiterpenes; C002781:ovalicin; D006639:Histidine; C026211:fumagillin; D000626:Aminopeptidases; C106541:methionine aminopeptidase 2; D008666:Metalloendopeptidases; D003545:Cysteine