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Protein & Peptide Letters

Volume 16 Issue 9
ISSN: 0929-8665
eISSN: 1875-5305


   All Titles

  Gβγ-Copurified Lipid Kinase Impurity from Sf9 Cells
  pp.1053-1056 (4) Authors: Aliaksei Shymanets, Mohammad Reza Ahmadian, Bernd Nurnberg

G-protein βγ dimers are prime regulators transmitting extracellular signals to wide-ranging cellular effectors including phosphoinositide 3-kinase (PI3K) isoforms β and γ. Recombinant Gβ?? purified from Sf9 cells via metal-affinity and anion exchange chromatography exhibited a wortmannin-insensitive phospholipid kinase activity that copurified from the insect cells. To exclude false-positive results of Gβγ-dependent lipid kinase activity, the elimination of insect phospholipid kinase from Gβγ protein samples is necessary to avoid interference with the intrinsic lipid kinase activity of PI3K isoforms in reconstitution experiments. Here we describe an improved procedure of Gβ1γ2 purification from cell membranes that separates the contaminating phospholipid kinase.

  Keywords: Gel filtration, G-proteins, Gβγ, phospholipid kinase, PI3Kγ, signal transduction, size exclusion chromatography, wortmannin
  Affiliation: Institut fur Experimentelle und Klinische Pharmakologie und Toxikologie, Klinikum der Eberhard-Karls-Universitat, Lothar-Meyer-Bau, Wilhelmstrasse 56, D-72074, Tubingen,Germany.
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