ライフサイエンスコーパス: CRABP

1 CRABP(II) expression was shown to be induced in the uter

2 CRABP-II thus facilitates the ligation of RAR and marked

3 CRABPs are members of the superfamily of lipid binding p

4 We designed and expressed a CRABP I mutant (A35C/T57C), in which a small-scale confo

5 ABP (msCRABP) demonstrates the presence of a CRABP in invertebrates.

6 o measured binding of FA to a retinoic acid (CRABP-I) and a retinol (CRBP-II) binding protein and we

7 in substrate channeling between CRABP-1 and CRABP-2 was also supported by isotope dilution experimen

8 production by uterine epithelial cells [and CRABP(II) expression] was also observed if the prepubert

9 etabolism between cells expressing CRABP and CRABP(II) and suggests CRABP(II) may participate in reti

10 ir isoelectric point showed both CRABP I and CRABP II to be present in the cerebellum and P19 cells,

11 lar lipid-binding protein termed CRABP-I and CRABP-II and that uses them as RA sensors.

12 retinoic acid-binding proteins (CRABP-I and CRABP-II), a nuclear retinoic acid receptor (RAR alpha),

13 retinoic acid binding proteins (CRABP-I and CRABP-II).

14 retinoic acid-binding proteins (CRABP-I and CRABP-II).

15 retinoic acid-binding proteins (CRABP-I and CRABP-II).

16 ng functional difference between CRABP-I and CRABP-II, and point at a novel mechanism by which the tr

17 retinoic acid binding proteins, CRABP-I and CRABP-II, and the purified heterocomplexes indicate that

18 retinoic acid-binding proteins, CRABP-I and CRABP-II, have been well characterized.

19 lative dissociation constant of CRABP-II and CRABP-I (Kd (CRABP-II)/Kd (CRABP-I)) was determined to b

20 he presence of this NLS in holo- but not apo-CRABP-II.

21 Additional observations demonstrate that apo-CRABP-II is associated with endoplasmic reticulum (ER),

22 nt samples, there was no association between CRABP-H mRNA expression level and APL cellular sensitivi

23 A difference in substrate channeling between CRABP-1 and CRABP-2 was also supported by isotope diluti

24 eal a striking functional difference between CRABP-I and CRABP-II, and point at a novel mechanism by

25 sm that involves direct interactions between CRABP-II and RAR.

26 "Channeling" of RA between CRABP-II and RAR markedly facilitates the formation of t

27 No correlation was seen between CRABP II or CRBP I mRNA levels and the levels of either

28 rding to their isoelectric point showed both CRABP I and CRABP II to be present in the cerebellum and

29 riptional activity of RA can be regulated by CRABP-II.

30 Substituting the corresponding CRABP-II residues onto CRABP-I conferred upon this prote

31 Unfolding of labeled tetra-Cys CRABP I is accompanied by enhancement of FlAsH fluoresce

32 onprone mutant (FlAsH-labeled P39A tetra-Cys CRABP I).

33 n Cooperative Oncology Group protocol E2491, CRABP-II mRNA was modestly increased from day 0 values i

34 with PMSG, cells previously shown to express CRABP(II) and confirmed here to continue to express it i

35 elium to synthesize RA as well as to express CRABP(II).

36 SG-treated rats (shown previously to express CRABP).

37 larly, injection of an adenovirus expressing CRABP-II into mammary carcinomas that spontaneously deve

38 retinoid metabolism between cells expressing CRABP and CRABP(II) and suggests CRABP(II) may participa

39 At no time did cells expressing CRABP exhibit significant retinoic acid synthesis.

40 raced to an aberrantly high intratumor FABP5/CRABP-II ratio.

41 l uteri [shown previously to be negative for CRABP(II)] or by smooth muscle and stromal cells taken f

42 earch for a biologically meaningful role for CRABP-II, we examined its effect on RA-induced growth in

43 from CRBP-1 and all-trans retinoic acid from CRABP-2, but not from CRABP-1.

44 ans retinoic acid from CRABP-2, but not from CRABP-1.

45 r movement of RA from CRABP-II, but not from CRABP-I, to RAR strongly depended on the concentration o

46 The data suggest that transfer of RA from CRABP-I to RAR involves dissociation of the ligand from

47 In contrast, movement of RA from CRABP-II to the receptor is facilitated by a mechanism t

48 The rate constant for movement of RA from CRABP-II, but not from CRABP-I, to RAR strongly depended

49 model revealed that the change stemmed from CRABP-I/CRABP-II substitution of three spatially aligned

50 r components involved in RA processing (e.g. CRABP).

51 in approximately heart > liver > intestine > CRABP > CRBP.

52 d is a proapoptotic agent in cells with high CRABP-II/FABP5 ratio, but it signals through PPARbeta/de

53 ee all-trans retinoids and holo-CRBP-1, holo-CRABP-1, and holo-CRABP-2.

54 For holo-CRBP-1 and holo-CRABP-2, the k(cat)/K(m) values either decreased 5-fold

55 oids and holo-CRBP-1, holo-CRABP-1, and holo-CRABP-2.

56 The k(cat)/K(m) value for holo-CRABP-1, however, decreased ~65-fold in comparison with

57 Analysis of retinoid transfer from holo-CRABPs to P450 27C1 suggests that the decrease in k(cat)

58 acid residues in CRABP-II to the homologous CRABP-I residues resulted in loss of the ability of CRAB

59 of cellular retinoic acid-binding protein I (CRABP I) in the RA signaling was investigated by examini

60 he cellular retinoic acid binding protein I (CRABP I) occurs via a flexible portal region, which func

61 an cellular retinoic acid-binding protein I (CRABP I) was mutated to incorporate in a surface-exposed

62 n, cellular retinoic acid-binding protein I (CRABP I), in the presence of an inert crowding agent (Fi

63 of cellular retinoic acid-binding protein I (CRABP I).

64 of cellular retinoic acid binding protein I (CRABP I).

65 evealed that the change stemmed from CRABP-I/CRABP-II substitution of three spatially aligned residue

66 lar retinoic acid-binding proteins I and II (CRABP-I and -II, respectively) are transport proteins fo

67 Cellular retinoic acid-binding protein II (CRABP-II) is an intracellular lipid-binding protein that

68 Cellular retinoic acid-binding protein II (CRABP-II) undergoes nuclear translocation upon binding o

69 f cellular retinoic acid binding protein-II (CRABP-II) has been invoked as an important mechanism of

70 ding protein cellular RA-binding protein-II (CRABP-II).

71 roteins cellular RA binding protein type II (CRABP-II) and fatty acid binding protein type 5 in adipo

72 cellular retinoic acid-binding protein (II) (CRABP(II)) may have a role in the movement of retinoic a

73 cellular retinoic-acid binding protein(II) [CRABP(II)] in the uterus of the rat.

74 ranslocation in response RA and thus impairs CRABP-II-mediated activation of RAR.

75 ed arginine residues (Arg-111 and Arg-131 in CRABP-I; Arg-111 and Arg-132 in CRABP-II) that interact

76 d Arg-131 in CRABP-I; Arg-111 and Arg-132 in CRABP-II) that interact with the carboxyl group of retin

77 In contrast, RA caused no change in CRABP(II) message level, even at times as late as 48 h a

78 ingly, these turns are on linked hairpins in CRABP I and represent the best-conserved turns in the iL

79 e peptides, encompassing turns III and IV in CRABP I, have a strong intrinsic bias to form native tur

80 ; range, 0.16-4.13) relative to the level in CRABP-H protein-expressing NB4 cells (arbitrarily set at

81 ely, converting these amino acid residues in CRABP-II to the homologous CRABP-I residues resulted in

82 , our data strongly imply that variations in CRABP-II expression and RA binding activity are not caus

83 The existence of an invertebrate CRABP has significant evolutionary implications, suggest

84 ween RARgamma and one of the CRABP isoforms (CRABP II) during the ligand transfer to the receptor.

85 iation constant of CRABP-II and CRABP-I (Kd (CRABP-II)/Kd (CRABP-I)) was determined to be 2-3, in clo

86 t of CRABP-II and CRABP-I (Kd (CRABP-II)/Kd (CRABP-I)) was determined to be 2-3, in close agreement w

87 constants of R111M and R132M (Kd (R111M)/Kd (CRABP-II) and Kd (R132M)/Kd(CRABP-II)) were determined t

88 (Kd (R111M)/Kd (CRABP-II) and Kd (R132M)/Kd(CRABP-II)) were determined to be 40-45 and 6-8, respecti

89 L28C CRABP mutants were generated, and the inserted cysteine

90 haracteristic of the promoter region of most CRABPs analyzed.

91 Analysis of the expression of mouse CRABP I from a cDNA expression plasmid in COS-1 cells re

92 lated structural homology with bovine/murine CRABP I shows msCRABP has a ligand binding pocket that c

93 Compared with bovine/murine CRABP I, the deduced amino acid sequence of msCRABP is 7

94 We previously showed that CRABP-II, but not CRABP-I, delivers RA to RAR through direct protein-prote

95 we show that expression of CRABP-II, but not CRABP-I, markedly enhanced RAR-mediated transcriptional

96 In the nucleus, CRABP-II directly binds to the nuclear receptor RAR to f

97 Arg-111 and Arg-132 of CRABP-II were replaced with methionine by site-directed

98 ty of RAR stems directly from the ability of CRABP-II to channel retinoic acid to the receptor.

99 residues resulted in loss of the ability of CRABP-II to interact with RAR and to augment the recepto

100 ion of this residue abolishes the ability of CRABP-II to undergo nuclear translocation in response RA

101 ith this finding, the RA binding activity of CRABP in APL cells from three pretreatment cases (range,

102 In contrast, the addition of CRABP I did not significantly affect the interaction of

103 To determine whether this appearance of CRABP(II) was dependent on the production of RA, both E2

104 NLS, mediates ligand-induced association of CRABP-II with importin alpha and is critical for nuclear

105 Moreover, the coexpression of CRABP I in CV-1 cells did not markedly inhibit or enhanc

106 brium dissociation constants of complexes of CRABP-I or CRABP-II with RA were found to differ by 2-fo

107 ition, the relative dissociation constant of CRABP-II and CRABP-I (Kd (CRABP-II)/Kd (CRABP-I)) was de

108 o RA binding is critical for dissociation of CRABP-II from ER and, consequently, for mobilization of

109 ore, we show that RA-induced dissociation of CRABP-II from the ER requires SUMOylation of K102.

110 R), and that RA triggers the dissociation of CRABP-II from this location.

111 re, the mechanisms underlying the effects of CRABP-II on the transcriptional activity of RAR and the

112 in narrowing the conformational ensemble of CRABP I during folding.

113 extract containing a 10-fold molar excess of CRABP I was incubated with RAR alpha extract in the pres

114 The ability of E2 to induce expression of CRABP(II) suggests that it can enhance the activity of R

115 as also been shown to increase expression of CRABP(II).

116 In turn, KLF2 induces the expression of CRABP-II and RARgamma, further potentiating inhibition o

117 they suggest that constitutive expression of CRABP-II could have a facilitative role in the response

118 Stable expression of CRABP-II in mammary carcinoma SC115 cells enabled activa

119 The observations show that expression of CRABP-II in preadipocytes is repressed by all three comp

120 Conversely, diminished expression of CRABP-II renders these cells retinoic acid resistant.

121 Here we show that expression of CRABP-II, but not CRABP-I, markedly enhanced RAR-mediate

122 05 does not alter the kinetics of folding of CRABP I, which indicates that the flexible loop containi

123 data unequivocally establish the function of CRABP-II in modulating the RAR-mediated biological activ

124 We show here that RA induces interactions of CRABP-II with the E2 SUMO ligase Ubc9 and triggers SUMOy

125 ificantly retarded the unfolding kinetics of CRABP I without influencing the urea dependence of the u

126 Photoaffinity labeling of CRABP-I with [(3)H]RA was light- and concentration-depen

127 re was no change from pretreatment levels of CRABP-II mRNA (median, 0.98) or, in three relapse cases

128 tography procedures to examine the levels of CRABP-II mRNA and RA binding activity in APL patient sam

129 The identity and the localization of CRABP I in the cytoplasm as well as the nuclei were also

130 e demonstrate further that overexpression of CRABP-II in MCF-7 mammary carcinoma cells dramatically e

131 Specifically, overexpression of CRABP-II, in the absence of RA, up-regulated the express

132 thesis and is augmented by overexpression of CRABP-II.

133 tween the electrostatic surface potential of CRABP-I and II revealed the presence of a sole region di

134 data demonstrate that the surface region of CRABP-II containing residues Gln75, Pro81, and Lys102 is

135 Here, we localize the region of CRABP-II that mediates the interactions of this protein

136 ifferentiation stems from down-regulation of CRABP-II.

137 ons demonstrate that permanent repression of CRABP-II in mature adipocytes is exerted by the master r

138 In agreement with the known role of CRABP-II in enhancing the transcriptional activity of RA

139 observations emphasize the important role of CRABP-II in regulating the transcriptional activity of R

140 The primary sequence of CRABP-II contains three putative SUMOylation sites, cent

141 acids comprising the ligand binding site of CRABP-I.

142 of crowding on the equilibrium stability of CRABP I was less than our experimental error (i.e., < or

143 effect of crowding on the denatured state of CRABP I by measuring side-chain accessibility using iodi

144 The ligand-controlled NLS "switch" of CRABP-II may represent a general mechanism for posttrans

145 eveal that the pro- and antiviral effects of CRABPs are mediated by modulation of LD abundance, where

146 on constants of the site-directed mutants of CRABPs.

147 ecific amino acids in the RA-binding site of CRABPs by photoaffinity labeling.

148 This work focuses on CRABP-II, a cytosolic protein that moves to the nucleus

149 sufficient for maximal tumor suppression on CRABP-II overexpression.

150 nt in the cerebellum and P19 cells, and only CRABP II to be present in the choroid plexus.

151 ing the corresponding CRABP-II residues onto CRABP-I conferred upon this protein the ability to chann

152 ciation constants of complexes of CRABP-I or CRABP-II with RA were found to differ by 2-fold.

153 quamous), the levels of nuclear receptors or CRABPs, and the response of the cells to the growth-inhi

154 anization of msCRABP is conserved with other CRABP family members and the larger LBP superfamily.

155 al of FlAsH on the tetra-Cys-containing P39A CRABP I is sensitive to whether this protein is native o

156 llular retinoic acid-binding protein I (P39A CRABP I), which forms inclusion bodies when expressed in

157 may slow closure of the beta-barrel in P39A CRABP I relative to the wild type, leaving it vulnerable

158 the aggregation-prone intermediates of P39A CRABP I contain predominantly beta-strands structured in

159 Photolabeled CRABP-I was hydrolyzed with endoproteinase Lys-C to yiel

160 cognate intracellular lipid-binding protein CRABP-II.

161 Cellular RA binding protein (CRABP II) colocalized with RALDH 1.

162 n of cellular retinoic acid-binding protein (CRABP) and cellular retinoic-acid binding protein(II) [C

163 n of cellular retinoic-acid-binding protein (CRABP) and cellular retinol-binding protein (CRBP), as w

164 the cellular retinoic acid-binding protein (CRABP) II, in this process.

165 n), ILBP (ileal fatty acid-binding protein), CRABP I (cellular retinoic acid-binding protein), and CR

166 to the two known acidic RA-binding proteins CRABP I and II, the cerebellum expressed a third RA-bind

167 ir respective cognate lipid-binding proteins CRABP-II and FABP5.

168 by the intracellular lipid binding proteins CRABP-II and FABP5.

169 for cellular retinoic acid-binding proteins (CRABP-I and CRABP-II), a nuclear retinoic acid receptor

170 and cellular retinoic acid binding proteins (CRABP-I and CRABP-II).

171 and cellular retinoic acid-binding proteins (CRABP-I and CRABP-II).

172 by cellular retinoic acid-binding proteins (CRABP-I and CRABP-II).

173 des cellular retinoic acid-binding proteins (CRABPs) and fatty acid binding proteins (FABPs).

174 us isoforms of cellular RA binding proteins (CRABPs) and RA receptor gamma (RARgamma).

175 Cellular retinoic acid-binding proteins (CRABPs) I and II were detected in one and three of the e

176 ding proteins [cellular RA binding proteins (CRABPs)-I and -II].

177 ate cellular retinoic acid binding proteins (CRABPs).

178 ges cellular retinoic acid binding proteins (CRABPs).

179 and cellular retinoic acid-binding proteins (CRABPs).

180 cytoplasmic retinoic acid binding proteins, CRABP-I and CRABP-II, and the purified heterocomplexes i

181 cytoplasmic retinoic acid-binding proteins, CRABP-I and CRABP-II, have been well characterized.

182 by two intracellular lipid-binding proteins-CRABP-II, which targets RA to RAR, and FABP5, which deli

183 known role in direct delivery of RA to RAR, CRABP-II may have an additional, RA-independent, functio

184 ment in the CRABP-II promoter and to repress CRABP-II expression.

185 residue) peptides corresponding to the seven CRABP I turns were analyzed by circular dichroism and NM

186 This discovery of a Manduca sexta CRABP (msCRABP) demonstrates the presence of a CRABP in

187 s analyses demonstrate that K102 is the sole CRABP-II residue to be SUMOylated in response to RA.

188 ficant evolutionary implications, suggesting CRABPs appeared during the evolution of the LBP superfam

189 expressing CRABP and CRABP(II) and suggests CRABP(II) may participate in retinoic acid production an

190 With this technique, CRABP I could also be detected in the HL60 cell line.

191 e intracellular lipid-binding protein termed CRABP-I and CRABP-II and that uses them as RA sensors.

192 These results demonstrate that CRABP I, while it might be important for RA homeostasis,

193 We found that CRABP-II mRNA in APL cells from pretreatment patients (n

194 Surprisingly, the data indicate that CRABP-II also displays proapoptotic activities on its ow

195 rlies RA resistance in tumors, indicate that CRABP-II functions as a tumor suppressor, and suggest th

196 We previously reported that CRABP-II enhances the transcriptional activity of RAR by

197 The observations reveal that CRABP-II plays a critical role in sensitizing tumors to

198 Recent studies revealed that CRABP-II functions by "channeling" RA to RAR, thereby en

199 The data further show that CRABP-II is a direct target gene for the glucocorticoid

200 We show that CRABP-II, a predominantly cytosolic protein, massively u

201 We previously showed that CRABP-II enhances the transcriptional activity of the nu

202 We previously showed that CRABP-II, but not CRABP-I, delivers RA to RAR through di

203 In summary, our study demonstrates that CRABPs serve as an on-off switch that modulates the effi

204 that, in the presence of retinoic acid, the CRABP-II-RAR complex is a short-lived intermediate.

205 adipocyte differentiation by activating the CRABP-II/RARgamma path in preadipose cells, thereby upre

206 s to neuronal progenitors is mediated by the CRABP-II/RAR path and that the FABP5/PPARbeta/delta path

207 ccompanied by a proportional increase in the CRABP I fraction.

208 ciate with a cognate response element in the CRABP-II promoter and to repress CRABP-II expression.

209 nding protein alpha-response elements in the CRABP-II promoter.

210 ncomitantly with a transient increase in the CRABP-II/FABP5 ratio at early stages of differentiation.

211 The unique properties of the CRABP I mutant described in this work can be used to ins

212 ty acid composition without induction of the CRABP II message.

213 interaction between RARgamma and one of the CRABP isoforms (CRABP II) during the ligand transfer to

214 E2 administration induced expression of the CRABP(II) gene in the uterus within 4 h, and this induct

215 inding site in the 5'-flanking region of the CRABP(II) gene was also required for this induction.

216 rmation about the ligand binding site of the CRABP-I molecule in solution.

217 tion of endogenous retinoic acid between the CRABPs and the nuclear receptors and thus affect retinoi

218 places [3H]-all-trans-retinoic acid from the CRABPs and increases retinoic acid occupancy of the hete

219 he mechanisms by which RA transfers from the CRABPs to RAR were thus investigated directly.

220 although it binds with high affinity to the CRABPs.

221 inoic acid binds with comparable affinity to CRABP-I and the heterocomplexes, but with approximately

222 with approximately 10-fold less affinity to CRABP-II.

223 Some of the 7-oxa-7,8-dihydro-RAs bind to CRABP and RARalpha.

224 ance of FA binding to FABP, binding of FA to CRABP-I was entropically driven.

225 -E-isomers of UAB retinoids bound tightly to CRABPs and RAR alpha, the binding affinity of the all-E-

226 differential expression patterns of the two CRABPs suggest that they serve distinct biological funct

227 ns of the aromatic residues of the wild-type CRABP-II and the two mutants were sequentially assigned

228 are highly similar to that of the wild-type CRABP-II.

229 While CRABP-II does not contain an NLS in its primary sequence

230 staining of the transfected COS-1 cells with CRABP I-specific antibody.