Mechanistic and metabolic inferences from the binding of substrate analogues and products to arginase.
Arginase is a binuclear Mn(2+) metalloenzyme that catalyzes the hydrolysis of L-arginine to L-ornithine and urea. X-ray crystal structures of arginase complexed to substrate analogues N(omega)-hydroxy-L-arginine and N(omega)-hydroxy-nor-L-arginine, as well as the products L-ornithine and urea, complete a set of structural "snapshots" along the reaction coordinate of arginase catalysis when interpreted along with the X-ray crystal structure of the arginase-transition-state analogue complex described in Kim et al. [Kim, N. N., Cox, J. D., Baggio, R. F., Emig, F. A., Mistry, S., Harper, S. L., Speicher, D. W., Morris, Jr., S. M., Ash, D. E., Traish, A. M., and Christianson, D. W. (2001) Biochemistry 40, 2678-2688]. Taken together, these structures render important insight on the structural determinants of tight binding inhibitors. Furthermore, we demonstrate for the first time the structural mechanistic link between arginase and NO synthase through their respective complexes with N(omega)-hydroxy-L-arginine. That N(omega)-hydroxy-L-arginine is a catalytic intermediate for NO synthase and an inhibitor of arginase reflects the reciprocal metabolic relationship between these two critical enzymes of L-arginine catabolism.
Roy and Diana Vagelos Laboratories, Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6323, USA.
J D
JD
Cox
E
E
Cama
D M
DM
Colleluori
S
S
Pethe
J L
JL
Boucher
D
D
Mansuy
D E
DE
Ash
D W
DW
Christianson
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D001119:Arginase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001120:Arginine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D001667:Binding, Competitive / Q000235:genetics; D002384:Catalysis; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000235:genetics; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D006639:Histidine / Q000235:genetics; D046911:Macromolecular Substances; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009952:Ornithine / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity / Q000235:genetics; D014508:Urea / Q000737:chemistry / Q000378:metabolism
D004791:Enzyme Inhibitors; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D006639:Histidine; C068309:N(omega)-hydroxyarginine; D014508:Urea; D001120:Arginine; D009952:Ornithine; D001119:Arginase; D003545:Cysteine