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. 2004 Jun;24(12):5369–5382. doi: 10.1128/MCB.24.12.5369-5382.2004

FIG. 3.

FIG. 3.

Identification of cellular proteins interacting with the tyrosine-phosphorylated ORF5. (Left) Glutathione-Sepharose beads containing GST (lane 1), GST-ORF5 (lane 2), or tyrosine-phosphorylated GST-ORF5 (GST-pORF5) (lane 3) were resolved by SDS-PAGE, followed by immunoblotting (IB) with an antiphosphotyrosine antibody (αpY). (Right) Glutathione-Sepharose beads containing GST (lane 1), GST-ORF5 (lane 2), or phosphorylated GST-ORF5 (GST-pORF5) (lane 3) were mixed with lysates of Jurkat T cells. ORF5 binding proteins were resolved by SDS-PAGE and visualized by Coomassie blue staining. Arrows indicate the PI3 kinase catalytic subunit (p110), PI3 kinase regulatory subunit (p85), SLP-76 (p75), Lck, and Fyn (p55). Numbers in parentheses indicate the frequency of peptides of each cellular protein obtained from mass spectrometry.