ライフサイエンスコーパス: neurabin

1 ly are the neuronal proteins spinophilin and neurabin.

2 sponses by a neural tissue-specific protein, neurabin.

3 GS4 interact with the overlapping regions of neurabin.

4 isoforms contributed to their recognition by neurabins.

5 d suggested that PP1 mediated I-2 binding to neurabins.

6 P1 inhibitor, also attenuated I-2 binding to neurabins.

7 ted RhoGEF), RGS3 and RGS12, spinophilin and neurabin-1, SRC homology 3 domain and multiple ankyrin r

8 es between the canonical PP1-binding motifs (neurabin, (457)KIKF(460); G(M)/R(GL), (65)RVSF(68)).

9 the plasma membrane, a process that requires neurabin (a neural tissue-specific protein).

10 ant peptides representing central regions of neurabins also preferentially bound PP1gamma1 and PP1alp

11 fractionation of rat brain demonstrates that neurabin and p70(S6k) both localize to the soluble fract

12 The mRNA of neurabin and p70(S6k) show striking colocalization in br

13 Neurabin and spinophilin are homologous protein phosphat

14 or subunits suggests that the PDZ domains of neurabin and spinophilin are important for targeting PP1

15 Neurabin and spinophilin are neuronal scaffolding protei

16 binding data also allowed us to classify the neurabin and spinophilin PDZ domains as the first identi

17 We report the structures of both the neurabin and spinophilin PDZ domains determined using bi

18 However, thus far, it is still unknown how neurabin and spinophilin themselves are targeted to thes

19 Neurabins are protein phosphatase-1 (PP1) targeting subu

20 493) proteins identified residues 473-479 of neurabin as containing a crucial PP1gamma1-selectivity d

21 Neurabin attenuated adenosine A(1) receptor (A1R) signal

22 n of a conserved PP1-binding motif abolished neurabin binding to both proteins.

23 Next, the adaptor protein NAB-1 (neurabin) binds to F-actin and recruits active zone prot

24 The central domain of both neurabins bound PP1 and I-2, and mutation of a conserved

25 p70(S6k) and neurabin coimmunoprecipitate from transfected HEK293 cel

26 Spinophilin and neurabin contain a single PDZ domain, a common protein-p

27 Deletion of Pro(464) and Ile(465) in neurabin (deltaPI) to equally space a conserved cluster

28 In addition, neurabin domains required for oligomerization are essent

29 1 selectively interacts with spinophilin and neurabin, F-actin-targeting proteins, whereas PP1beta se

30 Immunoprecipitation of spinophilin or neurabin from crude brain extracts selectively coprecipi

31 Neurabins from rat brain lysates were sedimented by I-2

32 We expressed wild-type and mutant neurabins fused to green fluorescent protein in Cos7, HE

33 Inactivation of the neurabin gene enhanced A1R signaling and promoted the pr

34 4 of G(M)/R(GL) fused to residues 457-493 of neurabin (GST-G(M)/Nb) selectively bound to and inhibite

35 sion proteins containing residues 146-493 of neurabin (GST-Nb-(146-493)) or residues 1-240 of G(M)/R(

36 ely interact with one neurabin molecule in a neurabin homo-oligomer to form a ternary complex, repres

37 Filamentous actin binding protein neurabin I (NrbI) targets protein phosphatase-1 (PP1) to

38 Phosphorylation of GST-neurabin I (residues 318-661) by PKA significantly reduc

39 Neurabin I also copurified with both the alpha and gamma

40 vity was confirmed by immunoprecipitation of neurabin I and II from brain extracts from wild type and

41 racts, including the actin-binding proteins, neurabin I and neurabin II.

42 Immunoprecipitation of neurabin I and neurabin II/spinophilin from rat brain ex

43 Although the F-actin-binding proteins neurabin I and spinophilin (neurabin II) also bind PP1,

44 Here we show that neurabin I binds protein phosphatase 1 (PP1) and inhibit

45 il and sterile alpha motif domains abolished neurabin I dimerization and induced filopodium extension

46 Immunoprecipitation of neurabin I established its association with PP1 and I-2

47 A glutathione S-transferase (GST)-neurabin I fusion protein (residues 318-661) containing

48 Neurabin I interacted with PP1alpha in an overlay assay,

49 Neurabin I is a brain-specific actin-binding protein.

50 n N-terminal F-actin-binding domain dictated neurabin I localization at actin cytoskeleton and promot

51 Immune complex assays showed that neurabin I recruited an active PP1 via a PP1-docking seq

52 tions that attenuated PP1 binding to I-2 and neurabin I suggested distinct and overlapping sites for

53 Neurabin I, a neuronal actin-binding protein, binds prot

54 d restored stress fibers in cells expressing neurabin I.

55 d the displacement of PP1 for recruitment to neurabin I.

56 ed protein kinase A (PKA) phosphorylation of neurabin I.

57 sights into the assembly and regulation of a neurabin I/PP1 complex that controls actin rearrangement

58 Finally, p70S6K was excluded from neurabin I/PP1 complexes and required the displacement o

59 Neurabin-I actin-binding domain bundled F-actin, promote

60 Furthermore, the expression of a neurabin-I polypeptide unable to bind PP1 delayed the ma

61 ociation of protein phosphatase-1 (PP1) with neurabin-I through a canonical KIXF motif inhibited filo

62 In contrast, dimerization of neurabin-I via C-terminal coiled-coil domains and associ

63 ted by a fragment of the PP1-binding protein neurabin-I, Nb-(146-493), because of the selective inhib

64 - and protein phosphatase-1-binding protein, neurabin-I, promotes filopodia in neurons and nonneurona

65 on, they impaired the ability of PP1 to bind neurabin-I, the neuronal regulatory subunit, and G(M), t

66 d hippocampal neurons showed that endogenous neurabin II and I-2 colocalized at actin-rich structures

67 n phosphatase 1 catalytic subunit (PP1) with neurabin II and inactivate PP1 and block mitosis during

68 on of the phosphorylated DCX and spinophilin/neurabin II from DCX-synthesizing glioma cells indicated

69 o a block of anchorage-independent growth as neurabin II is a synergistic inhibitor of anchorage-inde

70 Interaction between phosphorylated DCX and neurabin II may induce the association of the protein ph

71 binding proteins neurabin I and spinophilin (neurabin II) also bind PP1, their role in PP1 isoform se

72 g the actin-binding proteins, neurabin I and neurabin II.

73 Immunoprecipitation of neurabin I and neurabin II/spinophilin from rat brain extracts sediment

74 Site-directed mutagenesis of neurabin implicates its PDZ domain in the interaction wi

75 Cotransfection of neurabin in HEK293 cells activates p70(S6k) kinase activ

76 lective targeting of PP1gamma1 to F-actin by neurabins in intact cells requires both the canonical PP

77 erely attenuated PP1gamma1 interactions with neurabins in vitro and in cells and disrupted PP1gamma1

78 Recombinant spinophilin and neurabin interacted with endogenous PP1 and also with ea

79 Reduction of endogenous neurabin levels by interference RNA (RNAi)-mediated knoc

80 way of its PDZ domain, the neuronal-specific neurabin may target p70(S6k) to nerve terminals.

81 Our mechanistic analysis of neurabin-mediated regulation of A1R signaling in this st

82 , A1R and RGS4 each likely interact with one neurabin molecule in a neurabin homo-oligomer to form a

83 Here, we report the identification of neurabin (neural tissue-specific F-actin binding protein

84 Spinophilin (SPL) and neurabin (NRB) are structurally similar scaffolding prot

85 death in wild-type (WT) mice did not affect neurabin-null mice or WT mice cotreated with an RGS4 inh

86 Coexpression of neurabin or spinophilin with Lfc resulted in their clust

87 ed in brain, where it interacted with either neurabin or spinophilin.

88 We demonstrate the neurabin-p70(S6k) interaction by yeast two-hybrid analys

89 ctly and differentially bind spinophilin and neurabin PDZ domains.

90 t disrupting the functions of a cytoskeletal neurabin/PP1 complex enhanced filopodia and impaired sur

91 Spinophilin residues 427-470, or homologous neurabin residues 436-479, were sufficient to bind PP1 i

92 zed F-actin-binding proteins spinophilin and neurabin, respectively.

93 rid analysis using the coiled-coil domain of neurabin revealed an interaction with Lfc, a Rho GEF.

94 These data identify neurabin-RGS4 as a novel tissue-selective regulatory mec

95 Moreover, these findings implicate the A1R/neurabin/RGS4 complex as a valid therapeutic target for

96 ive effect achieved by disruption of the A1R/neurabin/RGS4 complex is elicited by the on-site and on-

97 ation are essential for formation of the A1R/neurabin/RGS4 ternary complex, as well as for stable loc

98 In the absence of PP1gamma1, both neurabins showed enhanced association with PP1alpha but

99 Thus, interaction between Lfc and neurabin/spinophilin selectively regulates Rho-dependent

100 Analysis of PP1 binding to chimeric neurabins suggested that sequences flanking a conserved

101 A) holoenzymes containing spinophilin and/or neurabin target specific neuronal PP1 isoforms, facilita

102 el PP1gamma1-selective interaction domain in neurabin that may allow for selective regulation and/or

103 me of the structural determinants in PP1 and neurabins that together contribute to preferential targe

104 M)/R(GL) severely compromised the ability of neurabin to bind and inhibit both isoforms but did not a

105 d the rank order of PP1 isoform selection by neurabins to be PP1gamma1 > PP1alpha > PP1beta.

106 h structures, consistent with the ability of neurabins to target the PP1.I-2 complex to actin cytoske

107 pinophilin: conserved as residues 473-479 in neurabin) to VKDYDTW severely attenuated PP1gamma1 inter

108 argeting proteins DARPP-32, spinophilin, and neurabin were also unchanged.

109 s efficiently coprecipitated spinophilin and neurabin, whereas PP1beta immunoprecipitation did not.

110 PP1 binding, facilitated PP1 recruitment by neurabins, which also targeted I-2 to polymerized F-acti

111 PP1-binding motif attenuated interactions of neurabins with both isoforms.

112 s not involved in the weaker interactions of neurabins with PP1beta.