Table 1. In vitro and in vivo PIP binding specificities of LBDs used in the PIPline collection.
| Domain | In vitro interaction | In vitro techniques | Kd | In vivo localizatio : yeast | In vivo localization animal cells | Evidences for in vivo binding (yeast/animal) | References |
|---|---|---|---|---|---|---|---|
| 1xFYVEHRS | PI3P | liposomes, fat blot, SPR, crystalography, monolayer penetration analysis | ≈0.5μM | endosome / vacuole | mostly cytosolic | Dependent on type III PI3K (VPS34), sensitive to PI3K inhibitor (e.g. Wm) | Burd et al., 1998; Gillooly et al., 2000; Misra and Hurley 2000; Sankaran et al., 2001; Raiborg et al., 2001; Stahelin et al., 2002; He et al., 2009 |
| 1xPXp40 | PI3P | liposomes, fat blot, SPR, ITC, crystalography, monolayer penetration analysis | ≈5μM | endosome / vacuole | early endosome | Dependent on type III PI3K (VPS34), sensitive to PI3K inhibitor (e.g. Wm) | Ago et al., 2001; Kanai et al., 2001; Bravo et al., 2001; Ellson et al., 2001; Zhan et al., 2002, Malkova et al. 2006 |
| 1xPHFAPP1 | PI4P | liposomes, fat blot, SPR, crystalography, monolayer penetration analysis, NMR spectroscopy | ≈18.6 μM | golgi | golgi | Dependent on PI4K (Pik1 in yeast, PI4K-IIIa in human), in vivo protection of PI4P, expression mask binding of anti-PI4P antibody, sensitive to PI4K inhibitor (e.g. PAO), sensitive to PI4-pptases (e.g. SAC1, Inp54) | Dowler et al., 2001; Levine et al., 2002; Godi et al. 2004; Balla et al., 2005; Hammond et al., 2009a, Lenoir et al., 2010; He at al., 2011 |
| 1xPHOSBP | PI4P | fat blot (this study), liposomes, SPR | ≈3.5μM | golgi | golgi | Dependent on PI4K (Pik1 in yeast, PI4K-IIIa in human), sensitive to PI4K inhibitor (e.g. PAO), sensitive to PI4-pptases (e.g. SAC1, Inp54) | Levine et al., 1998; Levine et al., 2002; Balla et al., 2005; Niu et al., 2013 |
| 1xPHPLC | PI(4,5)P2 | fat blot (this study), liposomes, SPR, crystalography, NMR spectroscopy | ≈1μM | mostly cytosolic | PM | localization sensitive to PLC inhibitor and activator sensitive to 5-pptases (e.g. INPP5B, OCRL), dependent on PI4P-5Kinase (e.g. mss4), expression mask binding to anti-PI(4,5)P2 antibody | Ferguson et al., 1995; Lemmon et al., 1995; Levine et al., 2002; Varnai et al., 2002; Tuzi et al., 2003; Yu et al., 2004; Szentpetery et al., 2005; Zoncu et al., 2007 Hammond et al., 2009a; Hammond et al., 2009b |
| 1xTUBBY-C | PI(4,5)P2, PI(3,4)P2, PI(3,4,5)P3 | fat blot (this study), liposomes, SPR, crystalography | nd | PM (this study) | PM | localization sensitive to PLC inhibitor and activator, sensitive to 5-pptases (e.g. INPP5B, OCRL) | Santagata et al., 2001; Quinn et al., 2008; Szentpetery et al., 2009; Hammond et al., 2012 |
Abbreviations: PI3K (PI3-kinase); PI4K (PI4-kinase); pptase (phosphatase); SPR (Surface Plasmon Resonance); ITC (Isothermal Titration Calorimetry), NMR (Nuclear Magnetic Resonance).