Abstract
The focal adhesion protein testin is a modular scaffold and tumour suppressor that consists of an N-terminal cysteine rich (CR) domain, a PET domain of unknown function and three C-terminal LIM domains. Testin has been proposed to have an open and a closed conformation based on the observation that its N-terminal half and C-terminal half directly interact. Here we extend the testin conformational model by demonstrating that testin can also form an antiparallel homodimer. In support of this extended model we determined that the testin region (amino acids 52–233) harbouring the PET domain interacts with the C-terminal LIM1-2 domains in vitro and in cells, and assign a critical role to tyrosine 288 in this interaction.
Highlights
Human testin (NP_056456.1), encoded by the TES gene (Unigene Hs.592286), is a modular scaffold protein consisting of a cysteine rich (CR) region, a central PET (Prickle, Espinas, Testin) domain and three LIM (Linl-1, Isl-1, Mec-3) domains together constituting the C-terminal half of testin (Fig 1A) [1,2,3]
In an extensive testin interactome study in HeLa cells we used a set of green fluorescent protein (GFP)-fused testin protein variants as bait to analyse, via affinity purification-mass spectrometry (AP-MS), the composition of the protein complexes these testin variants participate in
Testin peptides that could only originate from endogenous testin, were present in the tryptic digest of the co-precipitated complexes using the ΔPET-GFP and the LIM1-3-GFP variant (Fig 1A) as bait
Summary
Human testin (NP_056456.1), encoded by the TES gene (Unigene Hs.592286), is a modular scaffold protein consisting of a cysteine rich (CR) region, a central PET (Prickle, Espinas, Testin) domain and three LIM (Linl-1, Isl-1, Mec-3) domains together constituting the C-terminal half of testin (Fig 1A) [1,2,3]. A role for testin in p38-signalling has been suggested [9], previous reports mainly provide evidence that testin is a component of the actin cytoskeleton. It interacts with multiple actin cytoskeletal proteins (zyxin, α-actinin, mammalian enabled (MENA), vasodilator stimulated phosphoprotein (VASP), ena/VASP-like protein (Evl), spectrin, paxillin, talin and transforming growth factor beta-1-induced transcript 1 (Hic-5) and is present in actin-rich
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
