ライフサイエンスコーパス: PGHS-1

1 PGHS-1 and 2 are of particular interest because they are

2 PGHS-1 and PGHS-2 are conformational heterodimers, each

3 PGHS-1 and PGHS-2 protein levels did not reflect the cha

4 PGHS-1 expression was not altered by prednisone in eithe

5 PGHS-1 is considered a basal enzyme; PGHS-2 is associate

6 PGHS-1 is found constitutively expressed in most healthy

7 PGHS-1-dependent ( small middle dot)NO consumption also

8 protein prostaglandin H2 synthase isoform 1 (PGHS-1) is the target of the nonsteroidal antiinflammato

9 Prostaglandin H synthase isoform-1 (PGHS-1) cyclooxygenase activity has a cooperative respon

10 hacin-pretreated prostaglandin H synthase 1 (PGHS-1) were examined by low-temperature electron parama

11 e and inhibit prostaglandin H(2) synthase 1 (PGHS-1).

12 human prostaglandin endoperoxide synthase-1 (PGHS-1) and PGHS-2 and purified the recombinant enzymes

13 se activities of prostaglandin H synthase-1 (PGHS-1) both become irreversibly inactivated during reac

14 oxidase cycle of prostaglandin H synthase-1 (PGHS-1) can serve as the initial oxidant for arachidonic

15 dase activity of prostaglandin H synthase-1 (PGHS-1) have been reported to produce a large (2-7-fold)

16 Prostaglandin H synthase-1 (PGHS-1) is a bifunctional heme protein catalyzing both a

17 Prostaglandin H synthase-1 (PGHS-1) is a constitutively expressed key enzyme in the

18 Prostaglandin endoperoxide H synthase-1 (PGHS-1) is expressed constitutively in murine NIH 3T3 ce

19 ally consumed by prostaglandin H synthase-1 (PGHS-1) through acting as a reducing peroxidase substrat

20 ith prostaglandin-endoperoxide H synthase-1 (PGHS-1), the constitutive cyclooxygenase.

21 ovine prostaglandin endoperoxide synthase-1 (PGHS-1)/S- flurbiprofen complex suggests (a) that the ca

22 ine prostaglandin endoperoxide H synthase-1 (PGHS-1)/S-flurbiprofen complex suggest that the enzyme i

23 staglandin endoperoxide H synthases 1 and 2 (PGHS-1 and -2) are the major targets of nonsteroidal ant

24 Prostaglandin H synthase isoforms 1 and 2 (PGHS-1 and -2) catalyze the first two steps in the biosy

25 Prostaglandin H synthase isoforms 1 and 2 (PGHS-1 and -2) each have a peroxidase activity and also

26 staglandin endoperoxide H synthases-1 and 2 (PGHS-1 and PGHS-2; also cyclooxygenases-1 and 2, COX-1 a

27 taglandin endoperoxide H synthases-1 and -2 (PGHS-1 and -2) are the major targets of nonsteroidal ant

28 Prostaglandin H synthase-1 and -2 (PGHS-1 and -2) catalyze the committed step in prostaglan

29 taglandin endoperoxide H synthases-1 and -2 (PGHS-1 and -2) convert arachidonic acid to prostaglandin

30 to detect prostaglandin H synthase-1 and -2 (PGHS-1 and -2) in normal human hepatocytes and human hep

31 alysis by prostaglandin H synthase-1 and -2 (PGHS-1 and -2) requires activation of the normally laten

32 rostaglandin endoperoxide synthase-1 and -2 (PGHS-1 and -2, respectively).

33 taglandin endoperoxide H synthases-1 and -2 (PGHS-1 and PGHS-2), four short amphipathic helices near

34 f prostaglandin H synthase-1 and synthase-2 (PGHS-1 and PGHS-2) in the normal lung and in allergic lu

35 tes for the peroxidase activities of PGHS-2, PGHS-1, and glutathione peroxidase (GPx).

36 ent generation of a free radical at Tyr-385 (PGHS-1 numbering) in the cyclooxygenase active site; the

37 ylamino)propyl]-3-methyl-1-triazene (NOC-7), PGHS-1 enzyme activity was inhibited in the presence of

38 We conclude that (a) PGHS-1 is the predominant enzyme that biosynthesizes PGE

39 Both allergic PGHS-1(-/-) and PGHS-2(-/-) mice exhibited decreased bas

40 ory system compliance, whereas only allergic PGHS-1(-/-) mice showed increased baseline resistance an

41 HS-2 expression, leukoregulin fails to alter PGHS-1 levels in either orbital or dermal fibroblasts, s

42 Although PGHS-1 and -2 are similar biochemically, a number of stu

43 e show that inducible NO synthase (iNOS) and PGHS-1 co-localize in atherosclerotic lesions of ApoE(-/

44 erspective is that membrane proteins such as PGHS-1 and -2, which are located on the lumenal surface

45 hesizes PGE(2) in the normal mouse lung; (b) PGHS-1 and PGHS-2 products limit allergic lung inflammat

46 s on the promoter region activates the basal PGHS-1 gene transcription.

47 ter the beginning of a 10 min period of BCO, PGHS-1 mRNA was increased in fetal brainstem and hypotha

48 Both PGHS-1 and -2 were found on the lumenal surfaces of the

49 and localization seen with antisera to both PGHS-1 and PGHS-2 were similar but were not identical.

50 3-HPAA appeared to be converted by PGHS-1 in a similar manner; however, conversion was less

51 minants for efficient oxygenation of DHLA by PGHS-1, play similar roles in the oxygenation of EPA and

52 EPA significantly inhibits AA oxygenation by PGHS-1.

53 ne the evidence for independent signaling by PGHS-1 and PGHS-2, and the complex mechanisms for regula

54 al antibodies specific for the constitutive (PGHS-1) or inducible (PGHS-2) forms of the enzyme were u

55 The basis for the cooperative PGHS-1 behavior and for the difference in cooperativity

56 Cyanoferric PGHS-1 exhibited a nu(Fe)(-)(CN) line at 446 cm(-1) and

57 curring between the monomers comprising each PGHS-1 dimer, we analyzed structures of PGHS-1 crystalli

58 Within the nuclear envelope, PGHS-1 and -2 were present on both the inner and outer n

59 Two forms of PGHS exist, PGHS-1 (COX-1) and PGHS-2 (COX-2).

60 rs to be due to the constitutively expressed PGHS-1.

61 Ferric PGHS-1 has a predominant six-coordinate high-spin heme a

62 Ferrous PGHS-1 has a single species of five-coordinate high-spin

63 The ferrous PGHS-1 CO complex exhibited three important marker lines

64 For PGHS-1, the rate of conversion of Intermediate I to Inte

65 I formation (k1) was 2.3 x 10(7) M-1 s-1 for PGHS-1 and 2.5 x 10(7) M-1 s-1 for PGHS-2, indicating th

66 strate access channel with an R0 of 35 A for PGHS-1 with the PGHS-1 inhibitor and an R0 of 21 A for P

67 nts both a novel proaggregatory function for PGHS-1 and a regulated mechanism for vascular ( small mi

68 dicating that the photolabeling observed for PGHS-1 was not due to the presence of [125I]TID in the a

69 The rates of reactions for PGHS-1 reconstituted with MnPPIX were approximately an o

70 jor determinant of substrate specificity for PGHS-1 but not for PGHS-2.

71 clooxygenase is about 10-fold lower than for PGHS-1 cyclooxygenase, and this difference may contribut

72 ired lower GdmHCl levels for PGHS-2 than for PGHS-1.

73 ue for PGHS-2 was much greater than that for PGHS-1.

74 d in bronchoalveolar lavage (BAL) fluid from PGHS-1(-/-) mice, relative to wild-type or PGHS-2(-/-) m

75 rupting the prostaglandin G/H synthase genes PGHS-1 and-2.

76 ity and decreased affinity for heme of H386A PGHS-1 imply that His386 helps optimize heme binding.

77 AA to the cyclooxygenase site of ovine H386A PGHS-1 reopens the constriction in the cyclooxygenase si

78 Heme titrations indicated that H386A PGHS-1 binds heme less tightly than does native PGHS-1.

79 could be restored (10-30%) by treating H386A PGHS-1 with cyclooxygenase inhibitors or AA, but not wit

80 e residue Ser-529 to be active against human PGHS-1; the S529A mutant is resistant to inactivation by

81 V509I PGHS-2 mutants, like recombinant human PGHS-1, did not show time-dependent inhibition with any

82 The promoter of the human PGHS-1 gene lacks a TATA box, has a very GC-rich region,

83 In PGHS-1, a wide doublet tyrosyl radical (34-35 G) was for

84 In PGHS-1, cyclooxygenase inhibition by tyrosine nitration

85 e inhibitor to the heme moiety to be 20 A in PGHS-1 and 18 A in PGHS-2.

86 When His386 was substituted with alanine in PGHS-1, the mutant retained <2.5% of the native peroxida

87 n prostaglandin H synthase-2 (PGHS-2) and in PGHS-1 reconstituted with mangano protoporphyrin IX (MnP

88 hyperresponsiveness in wild-type mice and in PGHS-1(-/-) and PGHS-2(-/-) mice.

89 onding was detected for the heme-bound CO in PGHS-1.

90 tive site residue, that was not conserved in PGHS-1.

91 or abolished cyclooxygenase cooperativity in PGHS-1, MnPGHS-1, and MnPGHS-2.

92 apidly formed narrow singlet EPR (25-26 G in PGHS-1; 21 G in PGHS-2), and the same line shapes persis

93 istopathology) were significantly greater in PGHS-1(-/-) mice compared with PGHS-2(-/-) mice, and bot

94 the heme iron and the proximal histidine in PGHS-1.

95 to characterize tyrosyl radical kinetics in PGHS-1 and -2 reacted with ethyl hydrogen peroxide.

96 munohistochemistry revealed Tyr nitration in PGHS-1 in aortic lesions but markedly less in adjacent n

97 ees well with the open distal heme pocket in PGHS-1.

98 distinct from the initial tyrosyl radical in PGHS-1.

99 to either Ile (the corresponding residue in PGHS-1), Ala, Glu, or Lys was expressed by transient tra

100 peroxynitrite is a key intermediary step in PGHS-1 activation.

101 kinetics of the initial peroxidase steps in PGHS-1 and -2 to quantify mechanistic differences betwee

102 lic glutathione peroxidase in PGHS-2 than in PGHS-1.

103 Heme replacement increased PGHS-1 cyclooxygenase cooperativity and changed PGHS-2 c

104 However, other forms of NO(x)() inhibit PGHS-1.

105 The "constitutive" isoform, PGHS-1, is thought to have housekeeping functions, and t

106 the activity of prostaglandin H(2) isoforms (PGHS-1 and PGHS-2).

107 Proteolytic cleavage of labeled PGHS-1 at Arg277 with trypsin established that [125I]TID

108 Mice lacking PGHS-1 were similar to wild-type mice in all of these pa

109 the enzymes of arachidonic acid metabolism, PGHS-1 and 5-LO.

110 ins to determine which domains of microsomal PGHS-1 are subject to photolabeling.

111 ifferent nitrogen oxide derivatives modulate PGHS-1 activity.

112 Moreover, PGHS-1 protein cannot be detected in KAT-50.

113 Substitutions of Thr-383 (histidine in most PGHS-1) with histidine or aspartate decreased cyclooxyge

114 S-1 binds heme less tightly than does native PGHS-1.

115 e inhibitor, were 32, 67, and 7.1 for native PGHS-1, R120Q PGHS-1, and Y355F PGHS-1, respectively.

116 oM, respectively, versus 4 microM for native PGHS-1.

117 cupancy of the cyclooxygenase site of native PGHS-1 had no effect on peroxidase activity.

118 ifferent from values obtained for the native PGHS-1, suggesting that this residue is not importantly

119 ay account for the ability of PGHS-2 but not PGHS-1 to efficiently oxygenate AA in intact cells when

120 a) solubilized, partially purified ovine (o) PGHS-1; (b) membrane-associated, recombinant oPGHS-1; an

121 us, the results of our studies of Arg-120 of PGHS-1 and -2 imply that interactions involved in the bi

122 es coding for amino acids 1-139 and 1-136 of PGHS-1 and PGHS-2, respectively, which include the signa

123 Acetylation of Ser-530 of PGHS-1 by aspirin abolishes all oxygenase activity and t

124 ad no significant effect on the abundance of PGHS-1 mRNA in any brain region.

125 Acetylation of PGHS-1 by ASA, in contrast, inhibited both lipoxygenase

126 e markedly the kinetics and the chemistry of PGHS-1 peroxidase inactivation.

127 otting data showed a similar distribution of PGHS-1 and -2 in subcellular fractions, and product anal

128 to attribute the independent functioning of PGHS-1 and PGHS-2 to differences in their subcellular lo

129 His386 of PGHS-1 is not formally part of either active site, but l

130 The mechanism of inhibition of PGHS-1 by N-(carboxyheptyl)maleimide was investigated.

131 cubated with taxol; a selective inhibitor of PGHS-1 had no effect.

132 imidazoles (A) were tested as inhibitors of PGHS-1 and 5-LO and were found to be weak to inactive as

133 rements were used to analyze the kinetics of PGHS-1 peroxidase self-inactivation during reaction with

134 ith a glutamine increases the apparent Km of PGHS-1 for arachidonate by 1,000-fold.

135 Varying levels of PGHS-1 and PGHS-2-specific immunofluorescence were seen

136 nd dermal fibroblasts express high levels of PGHS-1 mRNA and protein, the other abundant form of cycl

137 olution data on the subcellular locations of PGHS-1 and -2, we employed immunoelectron microscopy wit

138 ssential for 3-nitrotyrosine modification of PGHS-1 was confirmed by the absence of 3-nitrotyrosine i

139 e peroxidase activity during the reaction of PGHS-1 with EtOOH or 15-HPETE correlated with oxyferryl

140 Reaction of PGHS-1 with peroxide forms Intermediate I, which has an

141 n of reducing cosubstrate during reaction of PGHS-1 with peroxide protected the peroxidase activity t

142 f peroxidase inactivation during reaction of PGHS-1 with peroxide.

143 clooxygenase inactivation during reaction of PGHS-1 with several hydroperoxides.

144 e inactivation were examined in reactions of PGHS-1 reconstituted with heme or mangano protoporphyrin

145 nces (e.g. negative allosteric regulation of PGHS-1 at low concentrations of arachidonate (500-1000 n

146 that cyclooxygenase active site residues of PGHS-1 fall into five functional categories as follows:

147 ic acid (DHLA) in the cyclooxygenase site of PGHS-1 and the effects of active site substitutions on t

148 which bind the cyclooxygenase active site of PGHS-1, prevented the labeling of the 38-kDa carboxyl-te

149 at peroxynitrite is an activating species of PGHS-1.

150 determine the stereochemical specificity of PGHS-1 toward 2-phenylpropionic acid inhibitors.

151 also influences the substrate specificity of PGHS-1; an S530T substitution causes 40- and 750-fold de

152 is a determinant of the stereospecificity of PGHS-1 toward inhibitors of the 2-phenylpropionic acid c

153 We have determined the structure of PGHS-1 at 3 angstrom resolution with arachidonic acid (A

154 each PGHS-1 dimer, we analyzed structures of PGHS-1 crystallized under five different conditions incl

155 The X-ray crystal structures of PGHS-1 in complex with the NSAIDs flurbiprofen and bromo

156 its overall, is distinctly less than that of PGHS-1.

157 The Km values of PGHS-1 and -2 for arachidonate are the same, and all but

158 tely reverses the action of acetaminophen on PGHS-1.

159 signaling with the largest effects being on PGHS-1 pathways involving PGD, PGE, and PGF.

160 -tetramethylphenylenediamine, and Trolox) on PGHS-1 cyclooxygenase velocity.

161 C deficient in hematopoietic PGD synthase or PGHS-1 in the presence or absence of the PGHS-2 inhibito

162 ferential peroxide activation of PGHS-2 over PGHS-1 seen in mammals was conserved in the fish enzymes

163 ling: residues 148, 348, 404, and 504 (ovine PGHS-1 numbering).

164 n the crystal structure of Co(3+)-heme ovine PGHS-1 complexed with arachidonic acid, 19 cyclooxygenas

165 s A-D) that include residues 74-122 in ovine PGHS-1 (oPGHS-1) and residues 59-108 in human PGHS-2 (hP

166 In ovine PGHS-1 crystallized in the absence of an NSAID, there is

167 ll folding, and of the active sites of ovine PGHS-1 and human PGHS-2 using denaturation with guanidin

168 Replacement of Arg-120 of ovine PGHS-1 with a glutamine increases the apparent Km of PGH

169 ay crystal structure of the complex of ovine PGHS-1 with AcSHA, confirms that the inhibitor elicits i

170 he putative membrane binding domain of ovine PGHS-1.

171 d site-directed mutagenesis to prepare ovine PGHS-1 mutants having modifications of Arg120 (R120K, R1

172 ecause acetaminophen inhibits purified ovine PGHS-1 and murine recombinant PGHS-2 equally.

173 he cyclooxygenase activity of purified ovine PGHS-1 in a time- and concentration-dependent manner sim

174 With purified ovine PGHS-1, ( small middle dot)NO consumption requires perox

175 Co(3+) protoporphyrin IX-reconstituted ovine PGHS-1 (Co(3+)-oPGHS-1) and compare the effects of activ

176 cular gland microsomes with [125I]TID, ovine PGHS-1 was one of the major photolabeled proteins.

177 At alkaline pH, PGHS-1 is converted to a second CO binding conformation

178 Cleavage of photolabeled PGHS-1 with endoproteinase Lys-C yielded a peptide conta

179 generating peroxynitrite, activates purified PGHS-1 and also stimulates PGE(2) production in arterial

180 Under optimal conditions purified PGHS-1 oxygenates EPA with only 10% of the efficiency of

181 -nitrotyrosine modification site in purified PGHS-1 exposed to peroxynitrite.

182 However, Q203V PGHS-1 and PGHS-2 mutants catalyzed heterolytic cleavage

183 ere 32, 67, and 7.1 for native PGHS-1, R120Q PGHS-1, and Y355F PGHS-1, respectively.

184 As observed previously with R120Q PGHS-1, flurbiprofen was an ineffective inhibitor of R12

185 Furthermore, hematin-reconstituted PGHS-1, which was rapidly inhibited by N-(carboxyheptyl)

186 e not COX substrates differentially regulate PGHS-1 versus PGHS-2.

187 1 by a series of NSAIDs and isozyme-specific PGHS-1 and PGHS-2 inhibitors were determined.

188 "basal" isoform of prostaglandin H synthase (PGHS-1).

189 ms of PGHS exist, a constitutive form termed PGHS-1 and an inducible form termed PGHS-2.

190 s the first and last steps more readily than PGHS-1.

191 phic studies have led to the hypothesis that PGHS-1 and -2 associate with only one face of the membra

192 biochemical support for the hypothesis that PGHS-1 and -2 do associate with membranes through a mono

193 The results indicate that PGHS-1 cyclooxygenase cooperativity originates in the fe

194 pt developed from the crystal structure that PGHS-1 binds to membranes via four amphipathic helices l

195 chemically, a number of studies suggest that PGHS-1 and PGHS-2 function independently to form prostan

196 bital or dermal fibroblasts, suggesting that PGHS-1 is not involved in cytokine-dependent prostanoid

197 lices present near the amino terminus of the PGHS-1 and PGHS-2 isozymes act as membrane anchors.

198 s coding for amino-terminal sequences of the PGHS-1 and PGHS-2 joined to the green fluorescent protei

199 volved in the constitutive expression of the PGHS-1 gene, we constructed a 2075-base pair fragment (-

200 plus IFN-gamma altered the expression of the PGHS-1 isoenzyme in RAW 264.7 cells.

201 annel with an R0 of 35 A for PGHS-1 with the PGHS-1 inhibitor and an R0 of 21 A for PGHS-2 with the P

202 group that is not covalently linked with the PGHS-1 protein.

203 s involved in the binding of arachidonate to PGHS-1 and -2 are quite different and that residues with

204 overall strength of arachidonate binding to PGHS-1.

205 In contrast to PGHS-1, only the formation kinetics of the PGHS-2 tyrosy

206 t for arachidonate binding to PGHS-2 than to PGHS-1.

207 ly to arachidonate binding to PGHS-2 than to PGHS-1.

208 Indomethacin-treated PGHS-1 and nimesulide-treated PGHS-2 rapidly formed narr

209 sorbance throughout the indomethacin-treated PGHS-1 reaction.

210 idase self-inactivation in inhibitor-treated PGHS-1 and MnPGHS-1 was characterized by stopped-flow sp

211 ion was about 0.3 s(-)1 in inhibitor-treated PGHS-1 and much slower in MnPGHS-1 (0.05 s(-)1); as with

212 tra during inactivation of inhibitor-treated PGHS-1 were similar to those observed with PGHS-1 but we

213 t the cyclooxygenase site (inhibitor-treated PGHS-1) thus can influence markedly the kinetics and the

214 red during inactivation of inhibitor-treated PGHS-1, producing iron chlorin and heme-protein adduct s

215 genase activities of recombinant brook trout PGHS-1 and -2 were characterized to test the generality

216 -2 to operate at low AA concentrations, when PGHS-1 is effectively latent.

217 ring the composition of the FA pool in which PGHS-1 functions.

218 uch slower in MnPGHS-1 (0.05 s(-)1); as with PGHS-1 itself, the peroxidase inactivation rates were in

219 y crystal structures of their complexes with PGHS-1.

220 smic isoform (cPGES) pairs functionally with PGHS-1, the cyclooxygenase that ordinarily exhibits cons

221 soform-selective inhibitors interacting with PGHS-1 and PGHS-2.

222 d PGHS-1 were similar to those observed with PGHS-1 but were rather distinct in MnPGHS-1; the kinetic

223 than the wide doublet radical observed with PGHS-1 reconstituted with heme.

224 = 0.19 microM) but is much less potent with PGHS-1 (IC(50) = 20 microM).

225 In contrast to the situation with PGHS-1 itself, significant amounts of heme degradation o

226 lly 1.5-2 times higher with PGHS-2 than with PGHS-1.

227 1 for native PGHS-1, R120Q PGHS-1, and Y355F PGHS-1, respectively.

228 chemical specificity observed with the Y355F PGHS-1 mutant suggests that Tyr355 is a determinant of t