| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 266, Issue 10, 6168-6173, 04, 1991
T Shimokawa and WL Smith
Prostaglandin endoperoxide (PGH) synthase has a single iron protoporphyrin
IX which is required for both the cyclooxygenase and peroxidase activities
of the enzyme. At room temperature, the heme iron is coordinated at the
axial position by an imidazole, and about 20% of the heme iron is
coordinated at the distal position by an imidazole. We have used
site-directed mutagenesis to investigate which histidine residues are
involved in PGH synthase catalysis and heme binding. Individual mutant
cDNAs for ovine PGH synthases were prepared with amino acid substitutions
at each of 13 conserved histidines. cos-1 cells were transfected with each
of these cDNAs, and the cyclooxygenase and peroxidase activities of the
resulting microsomal PGH synthases were measured. Mutant PGH synthases in
which His-207, His-309, or His- 388 was replaced with either glutamine or
alanine lacked both activities. Gln-386 and Ala-386 PGH synthase mutants
exhibited cyclooxygenase but not peroxidase activities. Other mutants
exhibited both activities at varying levels. Because binding of heme
renders native PGh synthase resistant to cleavage by trypsin, we examined
the effects of heme on the relative sensitivities of native, Ala-204, Ala-
207, Ala-309, Ala-386, and Ala-388 mutant PGH synthases to trypsin as a
measure of the heme-protein interaction. The Ala-309 PGh synthase mutant
was notably hypersensitive to tryptic cleavage, even in the presence of
exogenous heme; in contrast, the native enzyme and the other alanine
mutants exhibited similar, lower sensitivities toward trypsin and, except
for the Ala-386 mutant, were partially protected from trypsin cleavage by
heme. Preincubation of the native and each of the alanine mutant PGH
synthases, including the Ala-309 mutant, with indomethacin protected the
proteins from trypsin cleavage. Thus, all the mutant proteins retain
sufficient three-dimensional structure to bind cyclooxygenase inhibitors.
Our results suggest that His-309 is one of the heme ligands, probably the
axial ligand, of PGH synthase. Two other histidines, His-207 and His-388,
are essential for both PGH synthase activities suggesting that either
His-207 or His-388 can serve as the distal heme ligand; however, the
trypsin cleavage measurements imply that neither His-207 nor His-388 is
required for heme binding. This is consistent with the fact that only 20%
of the distal coordination position of the heme iron of PGH synthase is
occupied by an imidazole side chain.
Essential histidines of prostaglandin endoperoxide synthase. His-309 is involved in heme binding
Department of Biochemistry, Michigan State University, East Lansing 48824.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  K. Valmsen, W. E. Boeglin, R. Jarving, I. Jarving, K. Varvas, A. R. Brash, and N. Samel A Critical Role of Non-active Site Residues on Cyclooxygenase Helices 5 and 6 in the Control of Prostaglandin Stereochemistry at Carbon 15 J. Biol. Chem., September 21, 2007; 282(38): 28157 - 28163. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. Seibold, T. Ball, L. C. Hsi, D. A. Mills, R. D. Abeysinghe, R. Micielli, C. J. Rieke, R. I. Cukier, and W. L. Smith Histidine 386 and Its Role in Cyclooxygenase and Peroxidase Catalysis by Prostaglandin-endoperoxide H Synthases J. Biol. Chem., November 14, 2003; 278(46): 46163 - 46170. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Lajoie, J. Sirois, and M. Dore Induction of Cyclo-oxygenase-2 Expression in Naturally Occurring Gastric Ulcers J. Histochem. Cytochem., July 1, 2002; 50(7): 923 - 934. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Koeduka, K. Matsui, Y. Akakabe, and T. Kajiwara Catalytic Properties of Rice alpha -Oxygenase. A COMPARISON WITH MAMMALIAN PROSTAGLANDIN H SYNTHASES J. Biol. Chem., June 14, 2002; 277(25): 22648 - 22655. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Liu, M. Antaya, A. K. Goff, D. Boerboom, D. W. Silversides, J. G. Lussier, and J. Sirois Molecular Characterization of Bovine Prostaglandin G/H Synthase-2 and Regulation in Uterine Stromal Cells Biol Reprod, March 1, 2001; 64(3): 983 - 991. [Abstract] [Full Text]
|
|
|
|
 |
|
 M. Oshima, N. Murai, S. Kargman, M. Arguello, P. Luk, E. Kwong, M. M. Taketo, and J. F. Evans Chemoprevention of Intestinal Polyposis in the Apc{{Delta}}716 Mouse by Rofecoxib, a Specific Cyclooxygenase-2 Inhibitor Cancer Res., February 1, 2001; 61(4): 1733 - 1740. [Abstract] [Full Text]
|
|
|
|
|
|
A. G. Spencer, E. Thuresson, J. C. Otto, I. Song, T. Smith, D. L. DeWitt, R. M. Garavito, and W. L. Smith The Membrane Binding Domains of Prostaglandin Endoperoxide H Synthases 1 and 2. PEPTIDE MAPPING AND MUTATIONAL ANALYSIS J. Biol. Chem., November 12, 1999; 274(46): 32936 - 32942. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. J. Rieke, A. M. Mulichak, R. M. Garavito, and W. L. Smith The Role of Arginine 120 of Human Prostaglandin Endoperoxide H Synthase-2 in the Interaction with Fatty Acid Substrates and Inhibitors J. Biol. Chem., June 11, 1999; 274(24): 17109 - 17114. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. M. Taketo Cyclooxygenase-2 Inhibitors in Tumorigenesis (Part I) J Natl Cancer Inst, October 21, 1998; 90(20): 1529 - 1536. [Abstract] [PDF]
|
|
|
|
|
|
A. G. Spencer, J. W. Woods, T. Arakawa, I. I. Singer, and W. L. Smith Subcellular Localization of Prostaglandin Endoperoxide H Synthases-1 and -2 by Immunoelectron Microscopy J. Biol. Chem., April 17, 1998; 273(16): 9886 - 9893. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. C. Goodwin, M. R. Gunther, L. C. Hsi, B. C. Crews, T. E. Eling, R. P. Mason, and L. J. Marnett Nitric Oxide Trapping of Tyrosyl Radicals Generated during Prostaglandin Endoperoxide Synthase Turnover. DETECTION OF THE RADICAL DERIVATIVE OF TYROSINE 385 J. Biol. Chem., April 10, 1998; 273(15): 8903 - 8909. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Boerboom and J. Sirois Molecular Characterization of Equine Prostaglandin G/H Synthase-2 and Regulation of Its Messenger Ribonucleic Acid in Preovulatory Follicles Endocrinology, April 1, 1998; 139(4): 1662 - 1670. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Chen, M. N. Salafranca, and J. L. Mehta Cyclooxygenase inhibition decreases nitric oxide synthase activity in human platelets Am J Physiol Heart Circ Physiol, October 1, 1997; 273(4): H1854 - H1859. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. M. Landino, B. C. Crews, J. K. Gierse, S. D. Hauser, and L. J. Marnett Mutational Analysis of the Role of the Distal Histidine and Glutamine Residues of Prostaglandin-Endoperoxide Synthase-2 in Peroxidase Catalysis, Hydroperoxide Reduction, and Cyclooxygenase Activation J. Biol. Chem., August 22, 1997; 272(34): 21565 - 21574. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Bakovic and H. B. Dunford Reactions of Prostaglandin Endoperoxide Synthase and Its Compound I with Hydroperoxides J. Biol. Chem., January 26, 1996; 271(4): 2048 - 2056. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. K. Bhattacharyya, M. Lecomte, C. J. Rieke, R. M. Garavito, and W. L. Smith Involvement of Arginine 120, Glutamate 524, and Tyrosine 355 in the Binding of Arachidonate and 2-Phenylpropionic Acid Inhibitors to the Cyclooxygenase Active Site of Ovine Prostaglandin Endoperoxide H Synthase-1 J. Biol. Chem., January 26, 1996; 271(4): 2179 - 2184. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Mancini, D. Riendeau, J.-P. Falgueyret, P. J. Vickers, and G. P. O'Neill Arginine 120 of Prostaglandin G/H Synthase-1 Is Required for the Inhibition by Nonsteroidal Anti-inflammatory Drugs Containing a Carboxylic Acid Moiety J. Biol. Chem., December 8, 1995; 270(49): 29372 - 29377. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Laneuville, D. K. Breuer, N. Xu, Z. H. Huang, D. A. Gage, J. T. Watson, M. Lagarde, D. L. DeWitt, and W. L. Smith Fatty Acid Substrate Specificities of Human Prostaglandin-endoperoxide H Synthase-1 and -2 J. Biol. Chem., August 18, 1995; 270(33): 19330 - 19336. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. S. C. Tam, D. H. S. Lee, E. Y. Wang, D. G. Munroe, and C. Y. Lau Tepoxalin, a Novel Dual Inhibitor of the Prostaglandin-H Synthase Cyclooxygenase and Peroxidase Activities J. Biol. Chem., June 9, 1995; 270(23): 13948 - 13955. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. D. Thuresson, K. M. Lakkides, C. J. Rieke, Y. Sun, B. A. Wingerd, R. Micielli, A. M. Mulichak, M. G. Malkowski, R. M. Garavito, and W. L. Smith Prostaglandin Endoperoxide H Synthase-1. THE FUNCTIONS OF CYCLOOXYGENASE ACTIVE SITE RESIDUES IN THE BINDING, POSITIONING, AND OXYGENATION OF ARACHIDONIC ACID J. Biol. Chem., March 23, 2001; 276(13): 10347 - 10357. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
