Tethering, protein

Groves, J. T., Wiilfing, C. and Boxer, S. G. (1996) Electrical manipulation of glycan-phosphatidyl inositol-tethered proteins in planar supported bilayers. Biophys.J., 71, 2716-2723. 

The immobilization strategies are of particular interest. The authors reasoned that the use of aldehydes to tether proteins to the solid phase could be ideal for certain protein-protein interaction studies. Since many protein-lysine residues are available for coupling to aldehydes via Schiff s base, a number of spatial orientations are possible. Such random oriented attachments would permit exposure of various surfaces of a protein to the solution, and new protein-protein interactions would be potentially possible. 

Dirac-Svejstrup AB, Shorter J, Waters MG, Warren G. 2000. Phosphorylation of the vesicle-tethering protein pi 15 by a casein kinase II-like enzyme is required for Golgi reassembly from isolated mitotic fragments. J Cell Biol 150 475-487. 

A more efficient approach for preventing non-specific interactions between tethered proteins and the surface consists in the intercalation of a thick hydrophilic cushion between the end-attached affinity ligands and the... 

During development of each ommatidium, a protein called Boss (Bride of Sevenless) is expressed on the surface of the R8 cell. This membrane-tethered protein is the ligand for the Sev RTK on the surface of the neighboring R7 precursor cell, signaling it to develop into a photosensitive neuron (Figure 14-18a). In mutant flies that do not express a functional Boss protein or Sev RTK, Interaction between the Boss and Sev proteins cannot occur, and no R7 cells develop (Figure... 

Lesa, G. M., Seemann, J., Shorter, J., Vandekerckhove, J., and Warren, G. (2000). The amino-terminal domain of the golgi protein giantin interacts directly with the vesicle-tethering protein pll5. J. Biol. Chem. 275, 2831-2836. 

Fig. 5 Schematic of the yeast sterol protein interaction network based on biochemical data and results of split-ubiquitin membrane yeast two-hybrid assays. The tethering protein, ERG28, is highlighted in red (Adapted from [52])...

The main advantage of the simulations is that they provide the exact solution of the model system. The main disadvantage is that they are very demanding computationally and therefore in order to make systematic studies a compromise must be reached between the molecular detail in the model system and the number of studies that can be carried out. Furthermore, in some applications, for example protein adsorption on tethered polymer layers, the time scale for the process may be too long to be able to be simulated even with a simple model. However, combined effective potentials obtained from other theoretical approaches with, for example, BD simulations may lead to studies of the kinetic prop>erties of protein adsorption in tethered protein layers. As computational capabilities increase we may be able to reach more complex and detailed systems by the use of straightforward simulations. 

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