Purification and SAXS analysis of the integrin linked kinase, PINCH, parvin (IPP) heterotrimeric complex.

ABSTRACT: The heterotrimeric protein complex containing the integrin linked kinase (ILK), parvin, and PINCH proteins, termed the IPP complex, is an essential component of focal adhesions, where it interacts with many proteins to mediate signaling from integrin adhesion receptors. Here we conduct a biochemical and structural analysis of the minimal IPP complex, comprising full-length human ILK, the LIM1 domain of PINCH1, and the CH2 domain of α-parvin. We provide a detailed purification protocol for IPP and show that the purified IPP complex is stable and monodisperse in solution. Using small-angle X-ray scattering (SAXS), we also conduct the first structural characterization of IPP, which reveals an elongated shape with dimensions 120×60×40 Å. Flexibility analysis using the ensemble optimization met

SUBMITTER: Stiegler AL 

PROVIDER: S-EPMC3561323 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature