Analyse von Phosphatidylinositol (3,4)-Bisphosphat bindenden Proteinen

• Antragstellerin: Dr. Sonja Krugmann
• Fachliche Zuordnung: Biochemie
• Förderung: Förderung von 2001 bis 2003
• Projektkennung: Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 5315870

Several lipids have been shown to act as intracellular second messengers involved in the control of key pathways. Phosphatidylinositol (3,4)-bisphosphate, which is produced by phosphoinositide 3OH-konase is a likely candidate for a lipid second messenger. Using affinity chromatography, two novel FYFE domain-containing proteins, p44 and p80, were idenfified as phosphatidylinositol (3,4)-bisphosphate-binding proteins. I propose to analyse p44 and p80 functions in vivo and in vitro using cell biology and biochemistry. Initially, lipid binding specificities of recombinant proteins and contributions of their FYVE domains will be analysed in vitro using surface plasmon resonance technology. A potential catalytic function of p80, which contains a P loop motif, will bei addressed using standard assays. p44 and p80 binding proteins will bei isolated to provide further insights into their functions. Further studies will address the cellular functions of p44 and p80 in vivo using immunocytochemistry. Specific antibodies shall be raised to allow a comparison of the cellular distribution of tagged, overexpressed versus endogenous proteins. Preliminary results indicating a possible involvement of p80 in the secretory pathway shall be addressed in experiments employing appropriate pharmacological reagents and using p80 point mutants.

• DFG-Verfahren: Forschungsstipendien
• Internationaler Bezug: Großbritannien