Fig 1: Molecular interactions defined by co-immunoprecipitation (CO-IP). (A) Various NPXY-motif containing protein fragments pulling down PTB-less CCM2 isoforms (CCM2-107, CCM2-116, and CCM2-212). CCM2-201 serves as positive control. Interactions between cytoplasmic tails of ß-Integrins and CCM2 isoforms were confirmed by IP with GST beads pull-down. (B) Various NPXY-motif containing protein fragments pulled down CCM2 exons (6, 6A, and 6B) with GST beads. Full-length CCM2 PTB domain serves as positive control, while CCM2-1209 as negative control. (C) Various NPXY-motif containing protein fragments pulling down CCM2 exons (6, 6A, and 6B) with HIS beads. NPXY motifs containing protein fragments are cytoplasmic tails of ß-Integrins and CCM1 or PTB cores. CCM2-1209 (CCM2 C-terminal fragment, residues 303-444, no PTB domain), CCM1 fragments, K2 (1st NPXY motif), K5 (2nd NPXY motif), and mock control. Mock control is host cell lysate. MagneGST Glutathione Particles (Promega) for GST-tagged bait proteins and dynabeads (Invitrogen) for HIS-tagged bait proteins. All target proteins were labeled with radioactive with S35.
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