The ZASP-like motif in actinin-associated LIM protein is required for interaction with the α-actinin rod and for targeting to the muscle Z-line

Abstract

The Z-line is a specialized structure connecting adjacent sarcomeres in muscle cells. α-Actinin cross-links actin filaments in the Z-line. Several PDZ-LIM domain proteins localize to the Z-line and interact with α-actinin. Actinin-associated LIM protein (ALP), C-terminal LIM domain protein (CLP36), and Z band alternatively spliced PDZ-containing protein (ZASP) have a conserved region named the ZASP-like motif (ZM) between PDZ and LIM domains. To study the interactions and function of ALP we used purified recombinant proteins in surface plasmon resonance measurements. We show that ALP and α-actinin 2 have two interaction sites. The ZM motif was required for the interaction of ALP internal region with the α-actinin rod and for targeting of ALP to the Z-line. The PDZ domain of ALP bound to the C terminus of α-actinin. This is the first indication that the ZM motif would have a direct role in a protein-protein interaction. These results suggest that the two interaction sites of ALP would stabilize certain conformations of α-actinin 2 that would strengthen the Z-line integrity.