TY - JOUR
A2 - Yazawa, T.
A2 - Zhang, Y.
A2 - Zambrano, N.
AU - Korb, Doreen
AU - Tng, Priscilla Y.
AU - Milenkovic, Vladimir M.
AU - Reichhart, Nadine
AU - Strauss, Olaf
AU - Ritter, Oliver
AU - Fischer, Tobias
AU - Benz, Peter M.
AU - Schuh, Kai
PY - 2013
DA - 2013/02/07
TI - Identification of PDZ Domain Containing Proteins Interacting with 1.2 and PMCA4b
SP - 265182
VL - 2013
AB - PDZ (PSD-95/Disc large/Zonula occludens-1) protein interaction domains bind to cytoplasmic protein C-termini of transmembrane proteins. In order to identify new interaction partners of the voltage-gated L-type Ca2+ channel Cav1.2 and the plasma membrane Ca2+ ATPase 4b (PMCA4b), we used PDZ domain arrays probing for 124 PDZ domains. We confirmed this by GST pull-downs and immunoprecipitations. In PDZ arrays, strongest interactions with Cav1.2 and PMCA4b were found for the PDZ domains of SAP-102, MAST-205, MAGI-1, MAGI-2, MAGI-3, and ZO-1. We observed binding of the Cav1.2 C-terminus to PDZ domains of NHERF1/2, Mint-2, and CASK. PMCA4b was observed to interact with Mint-2 and its known interactions with Chapsyn-110 and CASK were confirmed. Furthermore, we validated interaction of Cav1.2 and PMCA4b with NHERF1/2, CASK, MAST-205 and MAGI-3 via immunoprecipitation. We also verified the interaction of Cav1.2 and nNOS and hypothesized that nNOS overexpression might reduce Ca2+ influx through Cav1.2. To address this, we measured Ca2+ currents in HEK 293 cells co-expressing Cav1.2 and nNOS and observed reduced voltage-dependent Cav1.2 activation. Taken together, we conclude that Cav1.2 and PMCA4b bind promiscuously to various PDZ domains, and that our data provides the basis for further investigation of the physiological consequences of these interactions.
SN - null
UR - https://doi.org/10.1155/2013/265182
DO - 10.1155/2013/265182
JF - ISRN Cell Biology
PB - Hindawi Publishing Corporation
KW -
ER -