TY - JOUR
T1 - PKB/Akt interacts with inosine-5' monophosphate dehydrogenase through its pleckstrin homology domain
AU - Ingley, Evan
AU - Hemmings, B.A.
PY - 2000
Y1 - 2000
N2 - The pleckstrin homology (PH) domain of the protooncogenic serine/threonine protein kinase PKB/Akt can bind phosphoinositides, A yeast-based two-hybrid system was employed which identified inosine-5' monophosphate dehydrogenase (IMPDH) type II as specifically interacting with PKB/Akts PH domain. IMPDH catalyzes the rate-limiting step of de novo guanosine-triphosphate (GTP) biosynthesis. Using purified fusion proteins, PKB/Akts PH domain and IMPDH associated in vitro and this association moderately activated IMPDH. Purified PKB/Akt also associated with IMPDH in vitro, We could specifically pull-down PKB/Akt or IMPDH from mammalian cell lysates using glutathione-S-transferase (GST)IMPDH or GST-PH domain fusion proteins, respectively, Additionally, PKB/Akt and IMPDH could be co-immunoprecipitated from COS cell lysates and active PKB/Akt could phosphorylate IMPDH in vitro. These results implicate PKB/Akt in the regulation of GTP biosynthesis through its interaction with IMPDH, which is involved in providing the GTP pool used by signal transducing G-proteins. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
AB - The pleckstrin homology (PH) domain of the protooncogenic serine/threonine protein kinase PKB/Akt can bind phosphoinositides, A yeast-based two-hybrid system was employed which identified inosine-5' monophosphate dehydrogenase (IMPDH) type II as specifically interacting with PKB/Akts PH domain. IMPDH catalyzes the rate-limiting step of de novo guanosine-triphosphate (GTP) biosynthesis. Using purified fusion proteins, PKB/Akts PH domain and IMPDH associated in vitro and this association moderately activated IMPDH. Purified PKB/Akt also associated with IMPDH in vitro, We could specifically pull-down PKB/Akt or IMPDH from mammalian cell lysates using glutathione-S-transferase (GST)IMPDH or GST-PH domain fusion proteins, respectively, Additionally, PKB/Akt and IMPDH could be co-immunoprecipitated from COS cell lysates and active PKB/Akt could phosphorylate IMPDH in vitro. These results implicate PKB/Akt in the regulation of GTP biosynthesis through its interaction with IMPDH, which is involved in providing the GTP pool used by signal transducing G-proteins. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
U2 - 10.1016/S0014-5793(00)01866-4
DO - 10.1016/S0014-5793(00)01866-4
M3 - Article
VL - 478
SP - 253
EP - 259
JO - Federation of European Biochemical Societies Letters
JF - Federation of European Biochemical Societies Letters
SN - 0014-5793
ER -