Tandem LIM domain-containing proteins, LIMK1 and LMO1, directly bind to force-bearing keratin intermediate filaments.

The cytoskeleton of the cell is constantly exposed to physical forces that regulate cellular functions. Selected members of the LIM (Lin-11, Isl-1, and Mec-3) domain-containing protein family accumulate along force-bearing actin fibers, with evidence supporting that the LIM domain is solely responsible for this force-induced interaction. However, LIM domain's force-induced interactions are not limited to actin. LIMK1 and LMO1, both containing only two tandem LIM domains, are recruited to force-bearing keratin fibers in epithelial cells. This unique recruitment is mediated by their LIM domains and regulated by the sequences outside the LIM domains. Based on in vitro reconstitution of this interaction, LIMK1 and LMO1 directly interact with stretched keratin 8/18 fibers. These results show that LIM domain's mechano-sensing abilities extend to the keratin cytoskeleton, highlighting the diverse role of LIM proteins in force-regulated signaling.

Spatial proximity of proteins surrounding zyxin under force-bearing conditions.

Sensing physical forces is a critical first step in mechano-transduction of cells. Zyxin, a LIM domain-containing protein, is recruited to force-bearing actin filaments and is thought to repair and strengthen them. Yet, the precise force-induced protein interactions surrounding zyxin remain unclear. Using BioID analysis, we identified proximal proteins surrounding zyxin under normal and force-bearing conditions by label-free mass spectrometry analysis. Under force-bearing conditions, increased biotinylation of α-actinin 1, α-actinin 4, and AFAP1 were detected, and these proteins accumulated along force-bearing actin fibers independently from zyxin, albeit at a lower intensity than zyxin. VASP also accumulated along force-bearing actin fibers in a zyxin-dependent manner, but the biotinylation of VASP remained constant regardless of force, supporting the model of a free zyxin-VASP complex in the cytoplasm being corecruited to tensed actin fibers. In addition, ARHGAP42, a RhoA GAP, was also identified as a proximal protein of zyxin and colocalized with zyxin along contractile actin bundles. The overexpression of ARHGAP42 reduced the rate of small wound closure, a zyxin-dependent process. These results demonstrate that the application of proximal biotinylation can resolve the proximity and composition of protein complexes as a function of force, which had not been possible with traditional biochemical analysis.

Synthesis and characterization of carbon nanofibers by catalytic chemical vapor deposition using non-ferromagnetic metal complexes.

Carbon nanofibers (CNFs) have wide applications in energy storage devices, electrically conducting composites, selective adsorbents, and catalyst supports. Catalytic chemical vapor deposition was carried out in this work to synthesize CNFs at mild temperatures of 700 and 800 degrees C. Non-ferromagnetic metal complexes of La, Nb, and Ti, spread on porous NaX-type zeolite support, were tested as new catalyst. CNFs ranging from 30 to 200 nm in diameter were obtained. Images of transmission electron microscopy showed encapsulated transition-metal nanoparticles by CNFs. X-ray diffraction patterns revealed the crystalline structures of La (FCC), Nb (BCC), and Ti (HCP) formed over zeolite. Magnetic hysteresis loops showed superconductivity from the CNF-encapsulated Nb at 2 K. Raman spectra showed that all the samples possessed graphitic and amorphous carbon structures. Based on the SEM images and Raman spectra, the three metals all catalyzed the synthesis of CNFs.