WD-repeat-propeller-FYVE protein, ProF, binds VAMP2 and protein kinase Czeta.
We have recently identified a protein, consisting of seven WD repeats, presumably forming a beta-propeller, and a domain identified in Fab1p, YOTB, VAC1p, and EEA1 (FYVE) domain, ProF. The FYVE domain targets the protein to vesicular membranes, while the WD repeats allow binding of the activated kinases Akt and protein kinase (PK)Czeta. Here, we describe the vesicle-associated membrane protein 2 (VAMP2) as interaction partner of ProF. The interaction is demonstrated with overexpressed and endogenous proteins in mammalian cells. ProF and VAMP2 partially colocalize on vesicular structures with PKCzeta and the proteins form a ternary complex. VAMP2 can be phosphorylated by activated PKCzeta in vitro and the presence of ProF increases the PKCzeta-dependent phosphorylation of VAMP2 in vitro. ProF is an adaptor protein that brings together a kinase with its substrate. VAMP2 is known to regulate docking and fusion of vesicles and to play a role in targeting vesicles to the plasma membrane. The complex may be involved in vesicle cycling in various secretory pathways.
Fritzius
Thorsten
T
Institute of Medical Virology, University of Zurich, Gloriastrasse 30, Zurich CH-8006, Switzerland.
Frey
Alexander D
AD
Schweneker
Marc
M
Mayer
Daniel
D
Moelling
Karin
K
D016475:3T3 Cells; D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000378:metabolism*; D002522:Chlorocebus aethiops; D006801:Humans; D047468:Immunoprecipitation; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D018613:Microscopy, Confocal; D010766:Phosphorylation; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism*; D013379:Substrate Specificity; D020798:Two-Hybrid System Techniques; D050988:Vesicle-Associated Membrane Protein 2 / Q000378:metabolism*; D000099221:Protein Kinase C zeta
D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D011493:Protein Kinase C; D050988:Vesicle-Associated Membrane Protein 2; D000099221:Protein Kinase C zeta; C494807:VAMP2 protein, human; C513679:WDFY2 protein, human