Phosphorylation of the 20-kDa regulatory light chain of myosin catalyzed by a Ca 2؉ /calmodulin-dependent myosin light chain kinase is important in the initiation of smooth muscle contraction and other contractile processes in non-muscle cells. It has been previously shown that residues 1-142 of smooth muscle myosin light chain kinase are necessary for high-affinity binding to actincontaining filaments in cells (1). To further localize the region of the kinase required for binding, a series of N-terminal deletion mutants as well as several N-terminal glutathione S-transferase fusion proteins were constructed. Cosedimentation assays showed that a peptide containing residues 1-75 binds to purified smooth muscle myofilaments. Furthermore, the N-terminal peptide was sufficient for high-affinity binding to actin stress fibers in smooth muscle cells in vivo. Alanine scanning mutagenesis in the fusion protein identified residues Asp-30, Phe-31, Arg-32, and Leu-35 as important for binding in vitro. There are two additional DFRXXL motifs located at residues 2-7 and 58 -63. The DFR residues in these three motifs were individually replaced by alanine residues in the full-length kinase. Each of these mutations significantly decreased myosin light chain kinase binding to myofilaments in vitro, and each abolished high-affinity binding to actin-containing filaments in smooth muscle cells in vivo. These results identify a unique structural motif comprised of three repeat consensus sequences in the N terminus of myosin light chain kinase necessary for high-affinity binding to actin-containing filaments.
Remote epitaxy (RE), substrate polarity can "penetrate" two-dimensional materials (2DMs) and act on the epi-layer, showing a prospective universal growth strategy. However, essentially, the role that 2DMs plays in RE has not been deeply investigated so far. Here, the RE of single-crystal films on the weakest polarity/iconicity substrate is realized to reveal its essence physical properties. Graphene facilitates attenuative charge transfer (ACT) from a substrate to epi-layer to construct remote interactions. Interfacial atoms are assembled into "incommensurate" epitaxial relationships through graphene to reduce misfit dislocations in the epilayer. Moreover, graphene reduces the atomic migration barrier, leading to a tendency toward a "layer-by-layer" growth mode. Such film growth mode is different with the conventional epitaxy (CE), and it is beneficial for the fast growth of epi-layers and the reduction of dislocations at coalescence boundaries. The insightful revelation of the role of graphene reveals the interface physics of RE and provides a more valuable guide to using 2DMs to expand three-dimensional materials (3DMs) for application in devices.
Wurtzite AlN is widely used for deep ultraviolet optoelectronic devices (DUV), which are generally grown along the [0001]-direction of the wurtzite structure on currently available substrates. However, huge internal electrostatic fields are presented within the material along [0001] axis induced by piezoelectric and spontaneous polarization, which has limited the internal quantum efficiency of AlN based DUV LEDs dramatically. The internal fields can be strongly reduced by changing the epitaxial growth direction from the conventional polar c-direction into less polar crystal directions. Twinned crystal is a crystal consisting of two or more domains with the same crystal lattice and composition but different crystal orientations. In other words, twins can be induced to change crystal directions. In this work we demonstrated that the epitaxial growth of () semi-polar AlN on (0001) AlN by constructing () and () twin structures. This new method is relative feasible than conventional methods and it has huge prospect to develop high-quality semi-polar AlN.
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