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With the aim of immobilizing enzymes in a defined orientation at the surface of LB bilayers, an original strategy has been previously developed in our group.
The diffusion zone has in general a defined orientation relative to the superalloy substrate and is built up by at least three phases embedded in a softer matrix, with significant differences in nanohardness.
While the full-length PLA2 binds to membranes with a defined orientation, an engineered PLA2 fragment ΔN10 that lacks the N-terminal ten residues binds to membranes with weaker affinity and at random orientation, and exhibits ∼100-fold lower enzymatic activity compared to the full-length PLA2, indicating the key role of the N terminus in PLA2 function.
Such an attachment of ribosomes to the IM may occur at discreet sites on the membrane to insure the entry of proteins in a defined orientation for proper assembly of membrane structures.
This motif is reminiscent of a cis-regulatory motif recently identified in the regulatory regions of Drosophila immune response genes which consists of a heterotypic pair of REL and GATA binding sites with a defined orientation relative to each other [48].
10.7554/eLife.01483.007 Figure 3. Ctrapsaps the TMD in a helical conformation with a defined orientation.
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Moreover, S-layer/streptavidin fusion proteins have been constructed in order to bind up to three biotinylated biomolecules per S-layer subunit in a highly defined orientation and position (Moll et al., 2002).
Furthermore, S-layer lattices as envelope structure covering the spherical containers (Fig. 4) constitute biomimetic 'artificial virus-like particles' enabling both stabilization of the nanocarriers and presenting addressor molecules in a highly defined orientation and special distribution (Sleytr et al., 2010, 2013).
It harbors a particular combination of cleavage sites that permits cloning of a nucleic acid in defined orientation and controlled labeling of only the desired fragments with fluorescent dyes (Figs. 1, 2, 3).
Such inorganic nanoparticles can be further used as anchor points to bind single proteins in a site-directed manner and defined orientation, which is necessary to mimic the ligand presentation in the cell membrane itself.
Then the sample was placed toward defined orientation in an unshielded room.
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