ライフサイエンスコーパス: AlF4-

1 AlF4- and S1.ADP-VO(4)3- complexes with specific states

2 ne 5'-O-(thiotriphosphate) (ATPgammaS) < ADP-AlF4 < ATP < ADP < ADP-vanadate.

3 6 +/- 0.3 A) and more significantly with ADP-AlF4 (-4.6 +/- 0.2 A).

4 plex with the transition state ATP mimic ADP.AlF4 (-) and compared them with the previous nucleic aci

5 ADP.BeFx is similar to S1*.ATP, while S1.ADP.AlF4- and S1.ADP.VO(4)3- are more similar to S1**.ADP.Pi

6 suggest that the conformations of the S1.ADP.AlF4-, S1.ADP.VO(4)3- and S1.ADP.BeFx complexes in the C

7 hange conformation upon formation of the ADP.AlF4- stabilized complex with the MoFe-protein.

8 Chi6a labeled at Cys347 also showed an AlF4--dependent increase in LY fluorescence (91%), indic

9 brain membranes were activated with GDP and AlF4- and deactivated in the presence of equal amounts o

10 from an alphai1-agarose column with GDP and AlF4-, and purified beta1gamma2HF dimer stimulates type

11 ion on the column was functional as GDP, and AlF4- specifically eluted alphaq from the column.

12 ce cholesterol enrichment inhibited PDGF and AlF4--induced MAP kinase activity (the major mechanism b

13 ant in maintaining both the active state and AlF4- in the guanine nucleotide binding pocket.

14 Because AlF4- complexes with GDP to stabilize an activated state

15 ing was impaired, suggesting that they bound AlF4- more poorly.

16 elease, resulting in decreased activation by AlF4- and increased thermolability.

17 and trypsin protection, since activation by AlF4- requires bound GDP.

18 tide binding (and hence normal activation by AlF4-) but is not important for receptor-mediated activa

19 ke alphas-R258A, has decreased activation by AlF4-, increased thermolability (both reversed in the pr

20 s an ordered conformation upon activation by AlF4-.

21 by phenylephrine to 18 +/- 5.5% at 5 min, by AlF4- to 26 +/- 8.4% at 20 min, and dose-dependently up

22 Activation of G proteins by AlF4-, NaF, or endothelin-1 resulted in redistribution o

23 nd, prepower stroke myosin heads (trapped by AlF4-).

24 y N,N'-p-phenylene dimaleimide) S1.ADP[C2H2].AlF4- and S1.ADP[C2H2].BeFx.

25 When Chi6b was labeled with LY on Cys210, AlF4- caused a 220% increase in LY fluorescence, indicat

26 Molecular dynamics simulations to the E1.AlF4 (-).ADP.3Na(+) structure indicated that 1) bound or

27 th the phosphate analogs aluminium fluoride (AlF4-), vanadate (VO(4)3-) and beryllium fluoride (BeFx)

28 activation by receptor or aluminum fluoride (AlF4-) and increased basal GDP release.

29 d the G protein activator aluminum fluoride (AlF4-), we determined if TGF alpha or AlF4- could induce

30 yllium fluoride (BeFx) or aluminum fluoride (AlF4-), yields stable complexes which mimic the intermed

31 e-treatment of cells with aluminum fluoride (AlF4-).

32 Together, GDP and fluoroaluminate (AlF4-) form a transition state analog of GTP that activa

33 ein RGS4 complexed with G(i alpha1)-Mg2+-GDP-AlF4 .

34 in-protein interactions between Gialpha1-GDP-AlF4- and the RGS domain or full-length RGS4 were detect

35 ffinity to the transition state of Gpa1 (GDP-AlF4--bound), and with much lower affinity to the inacti

36 ipitates GDP-bound G alpha h but not the GDP-AlF4--bound form.

37 s evidenced by its high affinity for the GDP-AlF4--bound forms of Goalpha and Gialpha and its relativ

38 nt alphaq, probably by binding to alphaq.GDP.AlF4-.

39 phosphate or the transition state analog GDP.AlF4-.

40 lexed with the transition-state analogue GDP.AlF4- at 1.65 A resolution.

41 -complexed transducin alpha-subunit, Gtalpha.AlF4-, but failed to elicit stimulation of Gtalpha GTPas

42 , inhibition of force at pCa 4.0 with 0.5 mM AlF4- decreased force to 0.04 +/- 0.01 of maximum (+/- S

43 2+-activated force was inhibited with 0.5 mM AlF4- or with 30 mM 2,3 butanedione-monoxime (BDM) durin

44 However, for both mutants, the ability of AlF4- to decrease the rate of GTPgammaS binding was impa

45 GDP (2 mM) was able to rescue the ability of AlF4- to protect the mutants, suggesting that they might

46 phosphate synthesis activated by addition of AlF4- to cells overexpressing recombinant alphaq, probab

47 as reduced even though the concentrations of AlF4- and Mg2+ were maintained.

48 titration with increasing concentrations of AlF4-, the 13C resonances demonstrated a marked loss of

49 ad been removed, excluding direct effects of AlF4- on the orientation of TnC in muscle fibers.

50 TER significantly sooner in the presence of AlF4- (similar times to Galphai2-transfected cells).

51 showed reduced protection in the presence of AlF4-.

52 for all three cell lines in the presence of AlF4-.

53 oride (AlF4-), we determined if TGF alpha or AlF4- could induce tyrosine phosphorylation of PKCdelta.

54 es that directly interact with either GDP or AlF4-.

55 ant to bind and be activated by GTPgammaS or AlF4- was assessed by trypsin protection assays.

56 ability in the presence of isoproterenol or AlF4- (a mixture of 10 microM AlCl3 and 10 mM NaF).

57 but was less efficient with isoproterenol or AlF4-.

58 alpha h upon activation with GTP gamma S or AlF4-.

59 The absence of Cl- did not prevent AlF4- binding to Gsalpha.

60 lting in enhanced thermolability and reduced AlF4--induced adenylyl cyclase stimulation and trypsin p

61 We have observed that in solution AlF4- did not cause Gs subunits to dissociate unless NaC

62 Aluminum tetrafluoride (AlF4-) activation of heterotrimeric G-protein alpha-subu

63 TPgammaS was a more effective activator than AlF4- for all mutants, with Gsalpha-E259D being the leas

64 We have concluded that AlF4-, a ligand which is capable of activating G protein

65 Unlike Ras, we find that AlF4- and BeF3- mediate complex formation between Cdc42H

66 he work of other investigators suggests that AlF4- causes subunit dissociation when it activates Gs.

67 ver, until recently it has been thought that AlF4- does not mediate effects on the Ras superfamily of

68 Additionally, the AlF4--induced interaction is weakened significantly by t

69 r-(123-140) retained its ability to bind the AlF4--complexed transducin alpha-subunit, Gtalpha.AlF4-,

70 We have characterized the AlF4--induced complex formation between the GDP-bound fo

71 For Gsalpha-E259D, the AlF4--induced activation defect was more pronounced at l

72 te that the apparent affinity of GAP for the AlF4--mediated complex is similar to the affinity observ

73 he (31)P NMR signals and, in the case of the AlF4- complex, slow hydrolysis of some of the excess ADP

74 ubunits, RGS16 preferentially recognized the AlF4--bound conformation of Gs alpha Asp229Ser.

75 herein, cells generated PGI2 in response to AlF4-, GTPgammaS, and ATP in a dose-dependent manner.

76 ichment enhanced PGI2 release in response to AlF4-, GTPgammaS, and ATP.

77 ctor activation, particularly in response to AlF4-.

78 d the gamma-phosphate of GTP (cholera toxin, AlF4- ion).

79 yrosine phosphorylation of PKCdelta, whereas AlF4- only slightly stimulated PKCdelta tyrosine phospho

80 Madin-Darby canine kidney (MDCK) cells with AlF4- (activator of heterotrimeric G protein alpha subun

81 Treatment of cells with AlF4- during the Ca2+ switch had little effect on the ki

82 Inhibition of force with AlF4- also had no effect when sTnC structure was monitor

83 fective unless the cells were incubated with AlF4-.

84 Preactivation of the protein with AlF4- before labeling led to a decreased incorporation o

85 ctrum of the ATP, ADP, ADP x BeFx, and ADP x AlF4- containing S complexes.

86 obtained with ADP, ATP ADP x BeFx and ADP x AlF4- were essentially identical in the presence of Co2+

87 ure at 2.0 A of rod transducin alpha x GDP x AlF4- in complex with the effector molecule PDEgamma and

88 lone and in complex with alpha(t/i1) x GDP x AlF4-.

89 ra of the S1 x MgADP x BeFx and S1 x MgADP x AlF4- complexes resemble to those obtained upon addition