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Free Content Gβ γ-Copurified Lipid Kinase Impurity from Sf9 Cells

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Abstract:

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: G-proteins; Gel filtration; Gβγ; PI3Kγ; phospholipid kinase; signal transduction; size exclusion chromatography; wortmannin

Document Type: Research Article

Publication date: 2009-09-01

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  • Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.
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