| 主权项 |
1. A process of designing a water-soluble transmembrane protein,
wherein the transmembrane protein is a G Protein-Coupled Receptor (GPCR), the method comprising: (1) generating a first library of putative water-soluble first modified transmembrane variants of a first transmembrane region of a native transmembrane protein, wherein each of said variants is generated by replacing a plurality of hydrophobic amino acids of the first transmembrane region, wherein the plurality of hydrophobic amino acids are selected from the group consisting of: Leucine (L), isoleucine (I), valine (V), and phenylalanine (F), and wherein each leucine, isoleucine, valine, and phenylalanine is replaced with a non-ionic polar amino acid selected from the group consisting of Q (or alternatively, N or S), T (or alternatively, N or S), T (or alternatively, N or S) and Y, respectively; (2) comparing structure scores and solubility scores of each said putative water-soluble first modified transmembrane variants in the first library and, preferably ranking the putative water-soluble first modified transmembrane variants using said structure scores and solubility scores in order to arrive at a second library of putative water-soluble first modified transmembrane variants, wherein said structure scores are obtained by scoring the propensity of forming alpha-helical structure of the first transmembrane regions of said variants, and wherein said solubility scores are obtained by scoring the water solubility prediction of the first transmembrane regions of said variants; (3) repeating steps (1) through (2) for a second, third, fourth, fifth, sixth, seventh or, preferably, all transmembrane regions of the protein; (4) identifying the amino acid and/or nucleic acid sequences of the water-soluble transmembrane protein comprising sequences which are not included in any transmembrane regions modified in steps (1) through (3), and including any extracellular or intracellular domain of the water-soluble transmembrane protein, thereby designing said water-soluble transmembrane protein. |
