ライフサイエンスコーパス: FBPase

1 FBPase accumulates to a higher level in the pep4 cell, s

2 FBPase also utilizes the endocytic pathway for transport

3 FBPase can be found on the vacuolar membrane and also at

4 FBPase degradation was inhibited in cells overexpressing

5 FBPase import into Vid vesicles requires the VID22 gene.

6 FBPase import is low in the glucose-starved cells and is

7 FBPase import requires ATP hydrolysis and is stimulated

8 FBPase is first imported to Vid (vacuole import and degr

9 FBPase is targeted from the cytosol to a novel type of v

10 FBPase is targeted from the cytosol to the vacuole for d

11 FBPase is targeted from the cytosol to the yeast vacuole

12 FBPase is targeted to the yeast vacuole for degradation

13 FBPase levels in the extracellular fraction decreased af

14 FBPase may be targeted to small vesicles before uptake b

15 FBPase sequestration into the vesicles is not affected i

16 FBPase sequestration into Vid vesicles required ATP and

17 FBPase trafficking to the vacuole involves two distinct

18 FBPase undergoes a quaternary transition from the canoni

19 FBPase was observed in areas close to the plasma membran

20 FBPases in such organisms may be components of metabolic

21 FBPases of heterotrophic organisms of distantly related

22 ssion of various chimeric mRNAs in LLC-PK(1)-FBPase(+) cells demonstrated that a single 8-base AU seq

23 We suggest that bifunctionality of PFK-2/FBPase-2 complements the metabolic zonation of the liver

24 -2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase-2).

25 nts block FBPase degradation by accumulating FBPase in the cytosol and also in small vesicles in the

26 postulated that monovalent cations activate FBPase by helping the Arg276 residue "deshield" the part

27 bacteria rapidly down-regulate an activated FBPase in order to avoid futile cycling?

28 However, VID22 affects FBPase import indirectly through a cytosolic factor.

29 -1 mutant, indicating that Vid24p acts after FBPase sequestration into the vesicles has occurred.

30 e that the Mtb genome encodes an alternative FBPase (GPM2, Rv3214) that can maintain gluconeogenesis

31 ture of the second member of this family, an FBPase/IMPase from Archaeoglobus fulgidus (AF2372), has

32 gism in eukaryotic FBPases from an ancestral FBPase having a central aqueous cavity and exhibiting sy

33 utated, FBPase degradation was defective and FBPase association with Vid vesicles was impaired.

34 High potency (IC50 = 16 nM) and FBPase specificity were achieved by linking a 2-aminothi

35 ailed to co-localize with actin patches, and FBPase in the extracellular fraction did not decrease as

36 of Ssa1p and Ssa2p in [URE3] propagation and FBPase degradation.

37 These new cargo proteins and FBPase interacted with the TORC1 complex during glucose

38 The Mtb genome contains only one annotated FBPase gene, glpX.

39 owed the same degradation characteristics as FBPase in that the short term starvation of cells led to

40 radation of gluconeogenesis enzymes, such as FBPase.

41 For this assay, FBPase was fused with a truncated form of alkaline phosp

42 Pases but large and hydrophilic in bacterial FBPases, similar to eFBPase.

43 tion, but regulatory mechanisms of bacterial FBPases are not well understood.

44 y disparate observations regarding bacterial FBPases, implicating a mechanism of feed-forward activat

45 erization of a series of orally bioavailable FBPase inhibitors identified following the combined disc

46 ved for 3 days, fructose-1,6-bisphosphatase (FBPase) and malate dehydrogenase 2 are degraded in the v

47 kers, including fructose-1,6-bisphosphatase (FBPase) and the vacuole import and degradation protein V

48 porcine liver fructose-1, 6-bisphosphatase (FBPase) are conserved residues and part of a loop for wh

49 neogenic enzyme fructose-1,6-bisphosphatase (FBPase) are in the extracellular fraction (periplasm).

50 --10 of porcine fructose-1,6-bisphosphatase (FBPase) are poorly ordered or are in different conformat

51 d inhibitors of fructose 1,6-bisphosphatase (FBPase) exhibited low oral bioavailability (OBAV) and th

52 a coli requires fructose-1,6-bisphosphatase (FBPase) for growth on gluconeogenic carbon sources.

53 c activation of fructose-1,6-bisphosphatase (FBPase) from Escherichia coli by phosphoenolpyruvate imp

54 Fructose-1,6-bisphosphatase (FBPase) governs a key step in gluconeogenesis, the conve

55 l structures of fructose-1,6-bisphosphatase (FBPase) has been implicated in regulatory and catalytic

56 ld-type porcine fructose-1,6-bisphosphatase (FBPase) has no tryptophan residues.

57 urine series of fructose-1,6-bisphosphatase (FBPase) inhibitors led to the discovery of a series of b

58 esign of potent fructose 1,6-bisphosphatase (FBPase) inhibitors that interact with the AMP binding si

59 te analogues as fructose 1,6-bisphosphatase (FBPase) inhibitors.

60 Liver fructose-1,6-bisphosphatase (FBPase) is a regulatory enzyme in gluconeogenesis that i

61 neogenic enzyme fructose-1,6-bisphosphatase (FBPase) is degraded in the vacuole when glucose is added

62 mbinant porcine fructose-1,6-bisphosphatase (FBPase) is explored by site-directed mutagenesis and kin

63 neogenic enzyme fructose-1,6-bisphosphatase (FBPase) is imported into Vid (vacuole import and degrada

64 neogenic enzyme fructose-1,6-bisphosphatase (FBPase) is induced when Saccharomyces cerevisiae are sta

65 neogenic enzyme fructose-1,6-bisphosphatase (FBPase) is subjected to catabolite inactivation and degr

66 Fructose-1,6-bisphosphatase (FBPase) is synthesized in yeast during glucose starvatio

67 Fructose-1,6-bisphosphatase (FBPase) is targeted to the vacuole for degradation when

68 neogenic enzyme fructose-1,6-bisphosphatase (FBPase) is targeted to Vid vesicles when glucose-starved

69 Fructose-1,6-bisphosphatase (FBPase) operates at a control point in mammalian glucone

70 t complexes of fructose 1, 6-bisphosphatase (FBPase) reveal competition between AMP and divalent cati

71 neogenic enzyme fructose-1,6-bisphosphatase (FBPase) to the vacuole for degradation.

72 (2-MC(GltA)) is fructose-1,6-bisphosphatase (FBPase), a key enzyme in gluconeogenesis.

73 ilar to that of fructose-1,6-bisphosphatase (FBPase), an enzyme involved in both CBBC and neoglucogen

74 kinase (PEPCK), fructose-1,6-bisphosphatase (FBPase), and glucose-6-phosphatase (G6Pase) gene transcr

75 better studied fructose-1,6-bisphosphatase (FBPase), in both cases from the moss Physcomitrella pate

76 eogenic enzyme, fructose-1,6-bisphosphatase (FBPase), in Saccharomyces cerevisiae.

77 nesis pathway, fructose-1, 6-bisphosphatase (FBPase), is induced when Saccharomyces cerevisiae are gr

78 eogenic enzyme, fructose-1,6-bisphosphatase (FBPase), is induced when Saccharomyces cerevisiae are st

79 eogenic enzyme, fructose-1,6-bisphosphatase (FBPase), is selectively targeted from the cytosol to the

80 enzymes such as fructose-1,6-bisphosphatase (FBPase), malate dehydrogenase, isocitrate lyase, and pho

81 Fructose-1,6-bisphosphatase (FBPase), the key enzyme in gluconeogenesis in the yeast

82 binding site of fructose 1,6-bisphosphatase (FBPase).

83 n porcine liver fructose-1,6-bisphosphatase (FBPase).

84 ad compounds of fructose-1,6-bisphosphatase (FBPase).

85 pathway such as fructose-1,6-bisphosphatase (FBPase).

86 se 6-phosphate by a fructose bisphosphatase (FBPase).

87 (PRK) and the fructose-1,6-bisphosphatases (FBPase) from pea and spinach.

88 vid mutants block FBPase degradation by accumulating FBPase in the cytosol

89 Both FBPase and malate dehydrogenase 2 were associated with a

90 strates in the reverse reaction catalyzed by FBPase.

91 phoenolpyruvate and sulfate activate E. coli FBPase by at least 300%.

92 here is the allosteric inhibition of E. coli FBPase by glucose 6-phosphate (Glc-6-P), the first metab

93 tent with that model, the complex of E. coli FBPase with Fru-2,6-P(2) remains in the R-state with dyn

94 nd AMP are synergistic inhibitors of E. coli FBPase, placing AMP/Glc-6-P inhibition in bacteria as a

95 id cycle, are shown here to activate E. coli FBPase.

96 isphosphatase (pFBPase) and Escherichia coli FBPase (eFBPase) differ in three respects.

97 nding-deficient mutants failed to complement FBPase import in Deltacpr1 and Deltavid22 mutants.

98 Under in vivo conditions, FBPase-delta60Pho8p was targeted to the vacuole via Vid

99 of liver GNG pathway intermediates confirmed FBPase as the site of action.

100 these intracellular structures that contain FBPase, the Vid vesicle marker Vid24p, and the endosomal

101 onstituted using Vid vesicles that contained FBPase-delta60Pho8p.

102 to cells that have been starved for 3 days, FBPase is degraded in the vacuole.

103 eletion of the UBC1 gene exhibited defective FBPase import.

104 tion pathway, mutants that failed to degrade FBPase in response to glucose were isolated using a colo

105 se media for longer periods of time degraded FBPase in the vacuole in response to glucose.

106 Vid vesicles then deliver FBPase to the vacuole for degradation.

107 cles and plays a critical role in delivering FBPase from the vesicles to the vacuole for degradation.

108 coneogenic carbon sources and has detectable FBPase activity.

109 6322), the first reported orally efficacious FBPase inhibitor.

110 tion 45 is small in all available eukaryotic FBPases but large and hydrophilic in bacterial FBPases,

111 on of AMP/Fru-2,6-P2 synergism in eukaryotic FBPases from an ancestral FBPase having a central aqueou

112 ucose-starved cells, levels of extracellular FBPase decrease rapidly.

113 ritical role in the decline of extracellular FBPase in response to glucose.

114 Moreover, the reduction of extracellular FBPase was also dependent on the VPS34 gene.

115 g that actin polymerization is important for FBPase degradation.

116 t phosphatase activity is also necessary for FBPase degradation.

117 7p phosphatase activity and are required for FBPase degradation.

118 tains an Armadillo (ARM) domain required for FBPase degradation.

119 However, Tor1p was dissociated from FBPase after the addition of glucose.

120 Furthermore, FBPase import was inhibited when cells overexpressed the

121 Furthermore, FBPase is associated with different forms of vesicles, w

122 to noncompetitive for K42T, I190T, and G191A FBPases.

123 expression of gluconeogenic enzymes (G6Pase, FBPase, and PCK1), thereby reversing CARM1-induced glyco

124 In the absence of this gene, FBPase and the Vid vesicle protein Vid24p associated wit

125 However, FBPase import required the ubiquitin-conjugating enzyme

126 Before vacuolar import, however, FBPase is sequestered inside a novel type of vesicle, th

127 ore it is delivered to the vacuole, however, FBPase is imported into intermediate carriers called Vid

128 bited high potency and specificity for human FBPase.

129 idazole phosphonic acid, 16, inhibited human FBPase (IC50 = 90 nM) 11-fold more potently than AMP and

130 series of benzimidazole analogues with human FBPase IC(50)s < 100 nM.

131 rent structural features compared with human FBPases, thereby offering a potential and species-specif

132 are not synergistic inhibitors of the Type I FBPase from Escherichia coli, and consistent with that m

133 the first structure of a prokaryotic Type I FBPase.

134 Conceivably, Type I FBPases from all eukaryotes may undergo similar global c

135 oscopy studies demonstrate that the imported FBPase is localized to the vacuole in the permeabilized

136 Mutants lacking Cpr1p were defective in FBPase import.

137 reduced Reg1p binding along with defects in FBPase degradation and Vid vesicle trafficking to the va

138 The defects in FBPase import seen for the Deltaubc1 and the K48R/K63R m

139 lic protein that mediates Vid22p function in FBPase import.

140 one of the ATP binding proteins involved in FBPase import.

141 es, we identified a requirement for SEC28 in FBPase degradation.

142 ally induce similar conformational states in FBPase.

143 encoding a single amino acid substitution in FBPase (S123F), which allowed a strain lacking a functio

144 ny heterotrophic bacteria, but are absent in FBPases of organisms that employ fructose 2,6-bisphospha

145 ral cavity and the evolution of synergism in FBPases.

146 hese non-nucleotide purine analogues inhibit FBPase in a similar manner and with similar potency as A

147 Monovalent cations inhibit FBPase either by distorting the geometry of the active s

148 16 also inhibited FBPase in primary rat hepatocytes and correspondingly re

149 s that affect actin polymerization inhibited FBPase degradation, suggesting that actin polymerization

150 us achieving proof-of-concept for inhibiting FBPase as a drug discovery target.

151 ased on its potent inhibition of human liver FBPase (IC(50) = 55 nM) and significant glucose lowering

152 This is the first study to identify liver FBPase as a previously unknown regulator of appetite and

153 udy was to determine the importance of liver FBPase in body weight regulation.

154 teric site comparable with that of mammalian FBPase.

155 ose 2,6-bisphosphate inhibition in mammalian FBPases.

156 for AMP/Fru-2,6-P(2) synergism in mammalian FBPases.

157 that of AMP, the natural inhibitor of murine FBPase (IC50 of 4.0 microM).

158 inities were decreased (3-22-fold) in mutant FBPases.

159 presence of FBPase antigen in these mutants, FBPase is completely inactivated in all vid mutants, ind

160 In some vid mutants, FBPase is found in punctate structures in the cytoplasm.

161 SEC28 and other coatomer genes were mutated, FBPase degradation was defective and FBPase association

162 We report the purification of novel FBPase-associated vesicles from wild-type cells to near

163 r the Mg2+ and K+ kinetics and activation of FBPase.

164 ls are fractionated, a substantial amount of FBPase is sedimentable in the high speed pellet, suggest

165 studies indicate that substantial amounts of FBPase were in the extracellular fraction (periplasm) du

166 The decline of FBPase in the extracellular fraction was dependent on th

167 l proline is required for the degradation of FBPase and MDH2 for both the vacuolar and non-vacuolar p

168 ed for the vacuolar-dependent degradation of FBPase and MDH2.

169 ective in the glucose-induced degradation of FBPase in the vacuole have been isolated.

170 lays an important role in the degradation of FBPase in the vacuole.

171 this study, we examined the distribution of FBPase at the ultrastructural level.

172 e the Mg2+-binding site to the AMP domain of FBPase.

173 py in probing the conformational dynamics of FBPase.

174 Constitutive expression of FBPase and fructose-6-phosphate-1-kinase coupled with th

175 oposed model may be relevant to all forms of FBPase, including the thioredoxin-regulated FBPase from

176 ngaged and T-state, loop-disengaged forms of FBPase.

177 Here, we reconstitute import of FBPase into isolated Vid vesicles.

178 onstitution of the glucose-induced import of FBPase into the vacuole in semi-intact yeast cells using

179 olic proteins are required for the import of FBPase into vesicles.

180 on and is directly involved in the import of FBPase into Vid vesicles.

181 Furthermore, the import of FBPase is a saturable process.

182 The import of FBPase was defined as the fraction of the FBPase that wa

183 Lack of growth due to inhibition of FBPase by 2-MC(GltA) was overcome by increasing the leve

184 nd the mechanism of allosteric inhibition of FBPase by AMP.

185 otomy and direct pharmacologic inhibition of FBPase in transgenic mice both returned food intake and

186 Inhibition of FBPase is considered a promising way to reduce hepatic g

187 ltA) was overcome by increasing the level of FBPase or by micromolar amounts of glucose in the medium

188 Furthermore, high levels of FBPase remained in the extracellular fraction in the Del

189 K experiments indicate that the majority of FBPase is sequestered inside the vesicles.

190 Despite the presence of FBPase antigen in these mutants, FBPase is completely in

191 ibited a significant decrease in the rate of FBPase degradation in vivo as compared with Deltassa1, D

192 Deletion of the first 10 residues of FBPase reduces k(cat) by 30-fold and Mg(2+) affinity by

193 tudies were performed to confirm the role of FBPase.

194 in growth conditions could alter the site of FBPase degradation.

195 e each been reported to be the major site of FBPase degradation.

196 , these results suggest that the AMP site of FBPase may represent a potential drug target for reducin

197 cids as AMP mimics targeting the AMP site of FBPase, which was achieved using a structure-guided drug

198 o bind to the allosteric AMP binding site of FBPase.

199 that in pairs form complete active sites of FBPase.

200 The X-ray structure of FBPase complexed with 5-aminoimidazole-4-carboxamide-1-b

201 A crystal structure of FBPase in a complex with three zinc cations and the prod

202 se to changes in the quaternary structure of FBPase, can account for the phenomena above.

203 hobic residues from two separate subunits of FBPase.

204 Thus, the glucose-induced targeting of FBPase into the vacuole can be reproduced in our in vitr

205 The glucose-induced targeting of FBPase to the vacuole for degradation occurs in cells gr

206 ith Glc7p was important for the transport of FBPase from intermediate vacuole import and degradation

207 copic studies demonstrate that the uptake of FBPase by the vacuole is mediated in part by an autophag

208 de the cell, and have deleterious effects on FBPase activity.

209 s that interact with the AMP binding site on FBPase despite their structural dissimilarity to AMP.

210 high specificity for the AMP binding site on FBPase.

211 TR, -290 mV for spinach PRK, -315 mV for pea FBPase, and -330 mV for spinach FBPase were obtained.

212 r thioredoxins f and m, for FTR, and for pea FBPase.

213 attributed to decreased expression of PEPCK, FBPase, and G6Pase due to increased acetylation of signa

214 -2-kinase/fructose-2,6-bisphosphatases (PFK2/FBPase), which modulate the intracellular concentration

215 d cells to the antimetabolic effects of PFK2/FBPase inhibition.

216 tures of mouse hepatocyes and kidney LLC-PK1-FBPase(+) cells.

217 responsive increase in PEPCK mRNA in LLC-PK1-FBPase+ cells is mediated by a p38 mitogen-activated pro

218 K signaling pathway, clonal lines of LLC-PK1-FBPase+ cells that express constitutively active (ca) an

219 is a global conformational change in porcine FBPase induced by Fru-2,6-P(2) in the absence of AMP.

220 ied at a subunit interface, which in porcine FBPase undergoes significant conformational change in re

221 Evidently in porcine FBPase, the actions of AMP at the allosteric site and Fr

222 ure, are in the canonical R-state of porcine FBPase but nevertheless retain sterically blocked AMP po

223 een the canonical R- and T-states of porcine FBPase.

224 The first potent FBPase inhibitors were identified using a structure-guid

225 he control of a mesophyll-specific promoter (FBPase::abi1-1, abbreviated to fa).

226 n semi-intact yeast cells using radiolabeled FBPase, an ATP regenerating system and cytosol.

227 When TORC1 was inactivated by rapamycin, FBPase degradation was inhibited.

228 structure (resolution, 1.45A) of recombinant FBPase from Escherichia coli, the first structure of a p

229 FBPase, including the thioredoxin-regulated FBPase from the chloroplast.

230 he vacuole, suggesting that Vid24p regulates FBPase targeting from the vesicles to the vacuole.

231 osol using an in vitro assay that reproduces FBPase import into Vid vesicles.

232 ore, the addition of purified Cpr1p restored FBPase import in both the Deltacpr1 and the Deltavid22 m

233 he findings suggest that potent and specific FBPase inhibitors represent a drug class with potential

234 ed with negative littermates, liver-specific FBPase transgenic mice had 50% less adiposity and ate 15

235 Spinach FBPase exhibited a more complicated behavior, with a sin

236 5 mV for pea FBPase, and -330 mV for spinach FBPase were obtained.

237 With the exception of spinach FBPase, titrations showed a single two-electron componen

238 defect; Deltassa2 cytosol did not stimulate FBPase import into import competent Vid vesicles, but wi

239 ion of purified recombinant Ssa2p stimulated FBPase import into Deltassa2 Vid vesicles, providing Del

240 id vesicles, but wild-type cytosol supported FBPase import into competent Deltassa2 vesicles.

241 Here, we demonstrated that FBPase was degraded outside the vacuole (most likely in

242 g, thereby providing the first evidence that FBPase inhibitors could improve glycemia in animal model

243 Kinetic studies indicate that FBPase association with these vesicles is stimulated by

244 Ultrastructural studies indicate that FBPase is in Vid/endosomes following glucose addition, s

245 We propose that FBPase is imported into these vesicles before entering t

246 Here, we show that FBPase import is independent of vacuole functions and pr

247 le in the high speed pellet, suggesting that FBPase is associated with intracellular structures in th

248 following glucose addition, suggesting that FBPase is internalized in response to glucose refeeding.

249 gher level in the pep4 cell, suggesting that FBPase is targeted to the vacuole for degradation.

250 The FBPase gene originated in bacteria in conjunction with t

251 esicle to vacuole-trafficking step along the FBPase degradation pathway.

252 sly unidentified metabolic redundancy at the FBPase-catalysed reaction step of the pathway.

253 r membrane in various mutants that block the FBPase degradation pathway.

254 To identify genes involved in the FBPase degradation pathway, mutants that failed to degra

255 To identify proteins involved in the FBPase degradation pathway, we cloned our first VID (vac

256 that these vesicles are intermediate in the FBPase degradation pathway.

257 suggest a new mechanism of regulation in the FBPase enzyme family: anionic ligands, most likely phosp

258 indicates the likelihood of synergism in the FBPase from Leptospira interrogans (lFBPase), and indeed

259 the first protein identified that marks the FBPase-containing vesicles and plays a critical role in

260 ene was identified by complementation of the FBPase degradation defect of the vid24-1 mutant.

261 of FBPase was defined as the fraction of the FBPase that was sequestered inside a membrane-sealed com

262 Vid24p is localized to the FBPase-containing vesicles as a peripheral membrane prot

263 4.13 are as equipotent as AMP with regard to FBPase inhibition.

264 energy balance in liver-specific transgenic FBPase mice and negative control littermates of both sex

265 g2+ affinity by 2-fold relative to wild-type FBPase.

266 respectively, in kcat relative to wild-type FBPase.

267 In the absence of functional Vid24p, FBPase accumulates in the vesicles and fails to move to

268 Whether FBPase functions only to regulate glucose or has other m

269 In this paper we investigated whether FBPase association with intracellular structures also ex

270 Evidently, the association of AMP with FBPase disorders loop 52-72, the consequence of which is

271 A exposure, particularly among carbon (xylA, FBPase, limEH, Chitinase, rgl, rgh, rgaE, mannanase, ara