ライフサイエンスコーパス: procaspase 9

1 he cytochrome c-induced binding of Apaf-1 to procaspase 9.

2 it serves as a cofactor in the activation of procaspase 9.

3 ctivating factor-1 (Apaf-1), and recombinant procaspase-9.

4 rial cytochrome c and attenuated cleavage of procaspase-9.

5 tem using recombinant APAF-1 and recombinant procaspase-9.

6 restores cytochrome c-mediated activation of procaspase-9.

7 in a ternary complex with cytochrome c, and procaspase-9.

8 s unable to associate with Apaf-1 or bind to procaspase-9.

9 hrome c, recombinant APAF-1, and recombinant procaspase-9.

10 le in apoptosis by binding to and activating procaspase-9.

11 by itself sufficient to recruit and activate procaspase-9.

12 ith cytochrome c, self-associate and bind to procaspase-9.

13 9 completely inhibit catalytic processing of procaspase-9.

14 en caspase recruitment domains of Apaf-1 and procaspase-9.

15 insulin on the TNF-alpha-induced cleavage of procaspase-9.

16 addition, XIAP was coimmunoprecipitated with procaspase-9.

17 lication of agents affecting the cleavage of procaspase-9.

18 ed interferon-gamma-induced S-nitrosation in procaspase-9.

19 apoptosis proteins) expression or levels of procaspase-9.

20 domain of TUCAN selectively binds itself and procaspase-9.

21 rization of Apaf-1 and thereby activation of procaspase-9.

22 Procaspases-9, -3, and -7 were processed in onconase-tre

23 caspase-9 activity, since overexpression of procaspase-9 accelerates the rate of apoptosis in active

24 cytosol, where it binds to Apaf-1 to form a procaspase-9-activating heptameric protein complex named

25 eport here the reconstitution of the de novo procaspase-9 activation pathway using highly purified cy

26 -9 by Apaf-1, we designed an in vitro Apaf-1-procaspase-9 activation system using recombinant compone

27 emical analysis of the reconstituted de novo procaspase-9 activation using highly purified cytochrome

28 Oligomerization-induced procaspase-9 activation, both within the apoptosome and

29 paf-1 complex and relieves the inhibition of procaspase-9 activation.

30 Procaspase-9 also synergistically promotes dATP binding

31 luding Bcl-2 (an inhibitor of apoptosis) and procaspase-9 (an effector of apoptosis) expression, and

32 gly, IL-12 promoted an altered processing of procaspase-9 and -3 and a decrease in the percentage of

33 In most cases, the mitochondrially localized procaspase-9 and -3 are released early during apoptosis

34 ochondrial membrane potential, processing of procaspase-9 and -3, and activation of caspase-3.

35 ced Bcl-2 : Bax interaction, and cleavage of procaspase-9 and -3.

36 sociation and accelerated the association of procaspase-9 and Apaf-1 in both intact cells and cell-fr

37 emical steps required for Apaf-1 to activate procaspase-9 and induce apoptosis.

38 that nitric oxide regulates the cleavage of procaspase-9 and its downstream proteins and, subsequent

39 -1 facilitates the proteolytic activation of procaspase-9 and maintains the hyperactive state of the

40 a the induction of Bcl-2), the inhibition of procaspase-9 and procaspase-3 activation, and the elevat

41 c events, including sequential activation of procaspase-9 and procaspase-3 and subsequent proteolysis

42 vitro, purified calpain cleaved recombinant procaspase-9 and procaspase-3 without activating either

43 of cytochrome c as well as the processing of procaspase-9 and procaspase-3.

44 paf-1-/- MEFs also induced the processing of procaspase-9 and procaspase-8, Bid cleavage, and apoptos

45 decreases the TNF-alpha-induced cleavage of procaspase-9 and subsequent apoptosis by regulating XIAP

46 TUCAN interferes with binding of Apaf1 to procaspase-9 and suppresses caspase activation induced b

47 , which is responsible for the activation of procaspase-9 and the maintenance of the enzymatic activi

48 rocaspase-3, procaspase-7, procaspase-8, and procaspase-9 and with cleavage of Bid and poly(ADP-ribos

49 effect of insulin on the coprecipitation of procaspase-9 and XIAP.

50 th sequential cleavage of procaspase 8, BID, procaspase 9, and procaspase 3.

51 bsequent Apaf-1 self-association, binding to procaspase-9, and formation of active Apaf-1 oligomers.

52 pression of apoptosome components Apaf-1 and procaspase-9, and limiting caspase-9 activity, since ove

53 , caspase-3 activities, and analyses of Bid, procaspase-9, and pro-caspase-3 cleavage.

54 ol agents, induced cleavage of procaspase-3, procaspase-9, and procaspase-8, along with the known cas

55 sing in cell extracts and in a reconstituted procaspase 9/Apaf1 apoptosome system.

56 y, when implementing the homodimerization of procaspase-9 as a prerequisite for activation, the calcu

57 We have found that cleavage of procaspase-9 at Asp(330) to generate p35, p10 or p37, p1

58 omes exposed after proteolytic processing of procaspase-9 at Asp315.

59 tinctive neoepitopes, exposed by cleavage of procaspase-9 at either Asp315 or Asp330, was co-localize

60 osome results in the proteolytic cleavage of procaspase-9 at the cleavage site PEPD(315) to yield the

61 Apaf-1 can form oligomers and may facilitate procaspase-9 autoactivation by oligomerizing its precurs

62 se-9 sets the overall duration of the timer, procaspase-9 autoprocessing activates the timer, and the

63 ribose) polymerase (PARP), procaspase 3, and procaspase 9; benzyloxycarbonyl-VAD, the pancaspase inhi

64 f caspase 8, expression of dominant negative procaspase 9 blocked procaspase 3 cleavage and the poten

65 unction of Apaf-1 is not only to oligomerize procaspase-9 but also to maintain the interaction of the

66 apoptosis was inhibited by dominant negative procaspase-9 but not by inhibition of caspase-8 activati

67 WD-40 repeats, associated with and activated procaspase-9, but failed to recruit procaspase-3 and ind

68 Activation of procaspase-9 by Apaf-1 in the cytochrome c/dATP-dependen

69 To elucidate the mechanism of activation of procaspase-9 by Apaf-1, we designed an in vitro Apaf-1-p

70 To elucidate the mechanism of activation of procaspase-9 by Apaf-1, we produced recombinant full-len

71 ondria and the subsequent transactivation of procaspase-9 by Apaf-1.

72 model has been applied for the activation of procaspase-9 by apoptotic protease-activating factor (Ap

73 We also show that procaspase-9 can recruit procaspase-3 to the Apaf-1-proc

74 m, so that unlike the effector procaspase-3, procaspase-9 cannot be processed by the apoptosome as a

75 e Apaf-1 CARD may be free to interact with a procaspase-9 CARD either before or during apoptosome ass

76 y the activation of procaspase-3 (cas-3) and procaspase-9 (cas-9).

77 tic cell death associated with activation of procaspase 9, caspase 3, and caspase 7.

78 emonstrate that XIAP functions downstream of procaspase-9 cleavage as an inhibitor of both proteolyti

79 ase-9 can recruit procaspase-3 to the Apaf-1-procaspase-9 complex.

80 n cells, M1 coimmunoprecipitated with Apaf-1-procaspase-9 complexes.

81 A procaspase-9 construct with a point mutation that abroga

82 hypothesis we produced recombinant forms of procaspase-9 containing mutations that disabled one or b

83 Procaspase-9 contains an NH2-terminal caspase-associated

84 Remarkably, however, procaspase-9 could also bind via its small subunit to th

85 s cerevisiae overexpressing human Apaf-1 and procaspase-9, critical components of the apoptosome, or

86 , which neutralizes Puma and Bim, or loss of procaspase 9 diminished OSI-027-induced apoptosis in vit

87 normal conditions and that denitrosation of procaspase-9 enhances its cleavage and consequent apopto

88 Combination treatment induced Apaf-1 and procaspase-9 expression in which cytochrome c releases b

89 by the Saveille reaction, immunoprecipitated procaspase 9 from SNAP-treated cells released 6-fold mor

90 itself and in complex with the prodomain of procaspase-9 have been determined at 1.6 and 2.5 A resol

91 e, we provide the first direct evidence that procaspase-9 homodimerizes within the apoptosome, marked

92 e with the additional WD-40 repeat activated procaspase 9 in vitro in response to cytochrome c and dA

93 or Apaf-1 prevents or reduces activation of procaspase-9 in a cell-free system.

94 The product of this Apaf-1 cDNA activated procaspase-9 in a cytochrome c and dATP/ATP-dependent ma

95 optotic signaling molecule that can activate procaspase-9 in a cytochrome c/dATP-dependent fashion.

96 trosation regulates activation of endogenous procaspase-9 in HT-29 cells.

97 he additional WD-40 repeat bind and activate procaspase-9 in response to cytochrome c and dATP.

98 -1-cytochrome c complex can bind and process procaspase-9 in the absence of additional dATP or cytoch

99 f procaspase-9 was substituted for wild-type procaspase-9 in the apoptosome.

100 s are deficient in their ability to activate procaspase-9 in the presence of cytochrome c and dATP wh

101 ulate S-nitrosation of an initiator caspase, procaspase-9, in a human colon adenocarcinoma cell line,

102 stitutively active and capable of processing procaspase-9 independent of cytochrome c an dATP.

103 eu enabled Apaf-1 to self-associate and bind procaspase-9 independent of cytochrome c, though still r

104 l injury initiates proteolytic processing of procaspase-9 into the large and small subunits, leading

105 at its mechanism of activation, like that of procaspase-9, involves association with an Apaf-1-relate

106 Despite the release of cytochrome c, procaspase 9 is not processed.

107 an untreated cells, confirming that cellular procaspase 9 is susceptible to nitrosylation.

108 In contrast, assuming a scenario in which procaspase-9 is activated allosterically upon binding to

109 one possible mechanism, we demonstrate that procaspase-9 is released from mitochondria in onconase-t

110 15 by an intrinsic autocatalytic activity of procaspase-9 itself.

111 st that the WD-40 repeats may be involved in procaspase-9-mediated procaspase-3 recruitment.

112 Theoretically, procaspase-9 might be activated in the mitochondria in a

113 n, we show here that aggregation of multiple procaspase-9 molecules can induce their activation indep

114 A procaspase-9 mutant (D315A) that cannot produce the p12

115 Apaf-1-mediated processing of procaspase-9 occurs at Asp-315 by an intrinsic autocatal

116 herein that apoptosome-mediated cleavage of procaspase-9 occurs exclusively through a CARD-displacem

117 esults show that S-nitrosation of endogenous procaspase-9 occurs in the HT-29 cells under normal cond

118 ation of the important interface residues in procaspase-9 or Apaf-1 prevents or reduces activation of

119 ase-3 but had no effect on the processing of procaspase-9 or the activity of prior activated caspase-

120 tive apoptosome is assembled from Apaf-1 and procaspase-9 (pc-9).

121 o its rapid autocatalytic cleavage, however, procaspase-9 per se contributed little to the activation

122 Indeed, procaspase-9 possessed higher affinity for the apoptosom

123 ia into the cytosol leading to activation of procaspase-9, procaspase-3 and PARP cleavage.

124 is pathway with purified recombinant Apaf-1, procaspase-9, procaspase-3, and cytochrome c from horse

125 factor-alpha (TNF-alpha)-induced cleavage of procaspase-9, procaspase-3, and poly-(ADP-ribose) polyme

126 l serine externalization, and proteolysis of procaspase-9, procaspase-3, gelsolin, and protein kinase

127 ivation pathway using purified ProT, Apaf-1, procaspase-9, procaspase-3, Hsp70, cytochrome c, PHAPI,

128 PIP2 inhibited procaspase 9 processing in cell extracts and in a recons

129 aspase recruitment domain (CARD) released by procaspase-9 processing activates nuclear factor kappaB

130 Like cIAP1, XIAP had no effect on procaspase-9 processing and was a more potent inhibitor

131 1 interacts with the apoptosome and prevents procaspase-9 processing as well as downstream procaspase

132 in HeLa cervical cancer cells, half-times of procaspase-9 processing, as well as the molecular timer

133 wed mitochondrial depolarization but blocked procaspase-9 processing, suggesting that M1 targeted the

134 f Apaf-1 and followed by a slower process of procaspase-9 recruitment and cleavage to form the p35/34

135 However, procaspase-9 recruitment and processing are accelerated

136 , thereby facilitating a continuous cycle of procaspase-9 recruitment/activation, processing, and rel

137 xpression in tissues and ability to activate procaspase-9 remain poorly characterized.

138 The protein level of Apaf-1, an activator of procaspase-9, remained constant with the application of

139 cytochrome c (cyt c) and dATP interacts with procaspase-9, resulting in the sequential cleavage and a

140 p53 infection, and ectopic overexpression of procaspase-9 sensitized cells to Ad-p53-induced apoptosi

141 , wherein the intracellular concentration of procaspase-9 sets the overall duration of the timer, pro

142 h Apaf-1 subunit is bound noncovalently to a procaspase-9 subunit via their respective CARD domains.

143 led us to reveal S-nitrosation of endogenous procaspase-9 that was immunoprecipitated from the HT-29

144 eoxycytidine (DAC) caused an accumulation of procaspase-9 through mRNA upregulation, but the cells di

145 ecule of Apaf-1 interacts with a molecule of procaspase-9 through two highly charged and complementar

146 The stoichiometric ratio of procaspase-9 to APAF-1 is approximately 1 to 1 in the co

147 Thus, our data suggest that modification of procaspase-9 to protect it from inappropriate cleavage a

148 Surprisingly, the absolute level of procaspase 9 was also elevated after 10 days of increase

149 Processing of procaspase 9 was not observed, and inhibition of CD95-de

150 se-3 activity when a point mutant (D330A) of procaspase-9 was substituted for wild-type procaspase-9

151 er conditions where Apaf-1 interactions with procaspase-9 were observed.

152 reduced the level of XIAP precipitated with procaspase-9, whereas insulin reversed this effect.

153 and that binds and suppresses activation of procaspase-9, which we term TUCAN (tumor-up-regulated CA