CH 2 sucre ), CH arom, p.277395057 ,
1H), Hz Hz 1H 1H 1H, vol.687, issue.8, pp.650685031573-1 ,
CH 2 sucre+benzyle ) ; 76,4 ; 77,5, CH sucre, vol.721, issue.142, pp.684-1286 ,
87 (s, 2H) (d, J = 9,0 Hz, 1H, H 1' ), Hz 1H 1H 1H 1H Hz, vol.415222841, issue.7d 2, pp.55648382637309-716 ,
CH 2 sucre+benzyle ), CH sucre, vol.7213, issue.138, pp.685-1284 ,
CH 2 sucre+benzyle ), CH sucre, vol.748, issue.141, pp.68-75 ,
CH 2 sucre+benzyle ), CH sucre, vol.73, issue.180, pp.684-12881 ,
57 (d, J = 12,5 Hz, 1H), Hz 1H 1H Hz 1H Hz 1H Hz 1H 1H 1H, vol.4165, issue.7, pp.210-44350647084936296 ,
CH 2 sucre+benzyle ), CH sucre, vol.76, issue.181, pp.67-76 ,
92 (s, 2H), Hz, 1H), pp.62-4857684, 2001. ,
CH 2 sucre+benzyle ), CH sucre, vol.74, issue.168, pp.68-72 ,
93 (s, 2H), pp.7885-691 ,
CH 2 sucre+benzyle ), CH sucreC arom, vol.72, issue.168, pp.683-12857 ,
44 (se, 1H, H 1' ), pp.4036-5920428, 2007. ,
40 (se, 1H, H 1' ), 1H, vol.5, issue.6, p.4092 ,
d, J = 9,5 Hz, 1H, H 1', 1H Hz 1H, vol.5, issue.8, pp.4667899509-1 ,
CH 3 acétate, pp.2044-2046 ,
t, J = 9,5 Hz, 1H, Hz, 1H Hz 1H 1H, vol.435, issue.7, pp.30-4295563235437, 2000. ,
57 (d, J = 3,5 Hz, 1H), Hz 1H Hz, vol.7, issue.7 2, pp.6979-6980 ,
CH 2 sucre ), CH sucreC arom, vol.8, issue.170, pp.194-169 ,
CH 3 acétate )CH 2 sucre ), CH sucre, vol.20, issue.148, pp.7-7 ,
ES) : calculée pour C 22 H 23 NNaO 11, 1163. ,
CH 3 acétate )CH 2 sucre ), CH sucre, vol.20, issue.152, pp.4-5 ,
CH 3 acétate )CH 2 sucre ), CH sucreC=O isatine, vol.20, issue.170, p.754 ,
dt, J 1 = 8,0 Hz, J 2 = 1,0 Hz, 1H, 1H 1H Hz 1H 1H, vol.586, issue.7, p.810512193440 ,
57 (s, 1H, NH), Hz, 1H), vol.7, issue.9, pp.314-315 ,
CH 2 sucre ), CH sucreC arom, vol.20, issue.170, pp.2070-2076 ,
19,9CH 3 acétate )CH 2 sucre ), CH sucre, vol.20, issue.145, p.6 ,
ES) : calculée pour C 30 H 27 N 3 NaO 13, 1442. ,
21 (dd, J 1 = 12,5 Hz, J 2 = 2,5 Hz, 1H), 1H, vol.4, issue.5, p.2728 ,
s, 1H, NH), 1H Hz 1H Hz, vol.7, issue.9d 2, pp.3-73652640339 ,
CH 3 acétate )CH 2 sucre ), CH sucre, vol.20, issue.1432, p.207 ,
(s, 1H) Le proton de la fonction acide carboxylique n'est pas visible sur le spectre, 1H 1H 1H, vol.7, issue.8, p.59830818 ,
Les protons de la fonction acide carboxylique et du groupement NH 2 ne sont pas visibles sur le spectre ,
100,0, p.3 ,
ES) calculée pour C 13 H 12 N 3, 1031. ,
s, 1H, NH, p.730388 ,
106,0, p.30 ,
une solution 10X Pour le préparer, il faut le diluer au 1/10 dans de l'eau MilliQ et ajuster son pH à 7,4 par addition de 5 mL de NaOH 1N Solution de résazurine La solution de travail de résazurine est préparée en ajoutant 2 mL de la solution stock (solution de résazurine dans le PBS-D à 1,25 mg/mL) à 98 mL de milieu MEM complet sans rouge de phénol et sans SVF ,
MEM (Eagle), GLUTAMAX I, Earle's salt supplémenté en, Sérum de veau foetal (SVF) 10% ,
?M ; 0,1% DMSO), additionné de 200 ng de GST-RTK cyt dans un tampon phosphate (20 mM de MOPS ; 10 mM de MgCl 2 ; 10 mM de MnCl 2 ; 1 mM de DTT ; 2,5 mM d'EGTA ; 10 mM de ?-glycérophosphate ; 1 mM de Na 3 VO 4 et 1 mM de NaF) est immobilisé sur plaque. Après 5 minutes d'incubation, 2 ?g de GST-PLC-? et 2 ?Ci ? 33 P-ATP en présence de 2 ?M d'ATP non radioactif sont additionnés, Après 1 heure d'incubation à 37°C, de l'EDTA (100 mM au final) est ajouté pour stopper la réaction. Les puits sont lavés au PBS-tween et les plaques sont lues avec un lecteur micro? Wallac ,
40 ?M) est mis à incuber à 30°C dans un mélange contenant 0,1 nM du domaine catalytique de PKA, 0,1 mM de ? 32 P-ATP, 2 mM de Mg(OAc) 2 , 70 ?M d'heptapeptide phosphoaccepteur «kemptide» dans 15 mM d'un milieu Hepes-NaOH (Ph 7,2) contenant 0,13 M de KCl, 0,3 mM d'EGTA, 0,3 mM d'EDTA et 1 mM de phosphate de sodium ,
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