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Titolo:
Expression, purification, characterization and homology modeling of activeAkt/PKB, a key enzyme involved in cell survival signaling
Autore:
Kumar, CC; Diao, R; Yin, ZZ; Liu, YH; Samatar, AA; Madison, V; Xiao, L;
Indirizzi:
Schering Plough Res Inst, Dept Tumor Biol, Kenilworth, NJ 07033 USA Schering Plough Res Inst Kenilworth NJ USA 07033 Kenilworth, NJ 07033 USA Schering Plough Res Inst, Dept Struct Chem, Kenilworth, NJ 07033 USA Schering Plough Res Inst Kenilworth NJ USA 07033 Kenilworth, NJ 07033 USA
Titolo Testata:
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
fascicolo: 3, volume: 1526, anno: 2001,
pagine: 257 - 268
SICI:
0304-4165(20010615)1526:3<257:EPCAHM>2.0.ZU;2-4
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN-KINASE-B; GLYCOGEN-SYNTHASE KINASE-3; STRUCTURE-BASED DESIGN; TUMOR-SUPPRESSOR; MOLECULAR-CLONING; PHOSPHATIDYLINOSITOL 3-KINASE; GLUCOSE-TRANSPORT; AKT PROTOONCOGENE; C-AKT; ACTIVATION;
Keywords:
akt1; expression; activation; kinetics; recombinant;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
53
Recensione:
Indirizzi per estratti:
Indirizzo: Kumar, CC Schering Plough Res Inst, Dept Tumor Biol, 2015 Galloping Hill Rd, Kenilworth, NJ 07033 USA Schering Plough Res Inst 2015 Galloping Hill RdKenilworth NJ USA 07033
Citazione:
C.C. Kumar et al., "Expression, purification, characterization and homology modeling of activeAkt/PKB, a key enzyme involved in cell survival signaling", BBA-GEN SUB, 1526(3), 2001, pp. 257-268

Abstract

Akt is a serine/threonine kinase that plays a critical role in cell survival signaling and its activation has been linked to tumorigenesis. Upregulation of Akt as well as its upstream regulator phosphatidylinositol-3 kinase (PI3K) has been found in many tumors and the negative regulator of this pathway PTEN/MMAC is a tumor suppressor. As a target for drug discovery, we have expressed and purified an active Akt1 enzyme from a recombinant baculovirus-infected Sf9 cell culture. Coexpression of Akt1 with the catalytic subunit of PI3K or treatment with okadaic acid during expression was found to generate an active enzyme in the insect cell culture system. We have optimized the kinase activity and developed a simple quantitative kinase assay using biotinylated peptide substrates. Using the purified active enzyme, we have characterized its physical, catalytic and kinetic properties. Since Akt is closely related to protein kinase C (PKC) and protein kinase A, the issue of obtaining selective inhibitors of this enzyme was addressed by comparison of the structures of catalytic domains of Akt and PKC, derived by homology modeling methods. A number of amino acid differences in the ATP bindingregions of these kinases were identified, suggesting that selective inhibitors of Akt can be discovered. However, the ATP binding regions are highly conserved in the three isoforms of Akt implying that the discovery of isoform-selective inhibitors would be very challenging. (C) 2001 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 06/04/20 alle ore 11:58:48