METHOD FOR DETERMINING protein conformation BY PROTEOLYSIS

摘要

1.method u043eu043fu0440u0435u0434u0435u043bu0435u043du0438u00a0 u0432u043bu0438u00a0u043du0438u00a0 the polymorphism or mutation of nucleic acid, the nucleic acid molecule, the structural properties of the protein, the encoded order anna nucleic acid molecule comprising (a) effect on the protein, the nucleic acid molecule u043au043eu0434u0438u0440u0443u0435u043cu044bu0439, several u043fu0440u043eu0442u0435u0430u0437u0430u043cu0438; and (b) u043eu043fu0440u0435u0434u0435u043b by whether the placeor to what extent is u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au043eu0435 fission; and, at the discretion of (with) the comparison of the u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au043eu0433u043e u0440u0430u0441u0449u0435u043fu043bu0435u043du0438u00a0 with u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au0438u043c u0440u0430u0441u0449u0435u043fu043b u0435u043du0438u0435u043c wild type protein is subjected to the same protease (the same u043fu0440u043eu0442u0435u0430u0437).;2.method of screening u0434u043bu00a0 u043eu043fu0440u0435u0434u0435u043bu0435u043du0438u00a0 options value of at least one gene, comprising (a) a model of the protein encoded by each of the u0443u043au0430u0437u0430u043du043d s options; (b) the impact of each protein, at least one u043fu0440u043eu0442u0435u0430u0437u043eu0439; (c) determining whether the placeor to what extent is u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au043eu0435 fission; and (d) a comparison of the u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au043eu0433u043e u0440u0430u0441u0449u0435u043fu043bu0435u043du0438u00a0 with u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au0438u043c disintegrating with protein d one type of under exposed to the same protease (the same u043fu0440u043eu0442u0435u0430u0437).;3. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 2, so that the protein are exposed to several u043fu0440u043eu0442u0435u0430u0437.;4. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 1 or 3, so that at least some of the u043fu0440u043eu0442u0435u0430u0437 attacking various sites within the protein.;5. method for 1 or 2, u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0, protease (protease) includes (include) one or more of the following enzymes: trypsin, u0445u0438u043cu043eu0442u0440u0438u043fu0441u0438u043d, proteinase k, u0430u043cu0438u043du043eu043fu0435u043fu0442 u0438u0434u0430u0437u0430, u043au0430u0440u0431u043eu043au0441u0438u043fu0435u043fu0442u0438u0434u0430u0437u0430, u044du043bu0430u0441u0442u0430u0437u0430, u043au0430u043bu043bu0438u043au0440u0435u0438u043d, u043cu0435u0442u0430u043bu043bu043eu044du043du0434u043eu043fu0435u043fu0442u0438u0434u0430u0437u0430, papain and pepsin.;6. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 1 or 2, so that several proteins are exposed to protease (u043fu0440u043eu0442u0435u0430u0437).;7. method for 1 or 2, u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 what proteins are exposed at the same time u043fu0440u043eu0442u0435u0430u0437, or vice versa.;8. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 1 or 2, so that the protein (proteins) are exposed to various u043fu0440u043eu0442u0435u0430u0437 either simultaneously or alternately.;9. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 1 or 2, so that the protein (proteins) are exposed to protease (u043fu0440u043eu0442u0435u0430u0437) in u0443u0441u043bu043eu0432u0438u00a0u0445 to protease activity (u043fu0440u043eu0442u0435u0430u0437).;10. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 1 or 2, so that the split protein (protein) to terminate their addition to the reaction mixture of at least one protease inhibitor.;11. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 1 or 2, so that the degree of u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au043eu0433u043e u0440u0430u0441u0449u0435u043fu043bu0435u043du0438u00a0 u043eu043fu0440u0435u0434u0435u043bu00a0u044eu0442 by u043eu0431u0449u0435u043fu0440u0438u043du00a0u0442u043eu0433u043e method for protein analysis.;12. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 p.11, however, that the analysis includes analysis by gel electrophoresis in the presence of u0434u043eu0434u0435u0446u0438u043bu0441u0443u043bu044cu0444u0430u0442u0430 gehle u043du0430u0442u0440u0438u00a0 (sds - page).;13. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 p.12, so that, after the analysis of u043fu0440u043eu0432u043eu0434u00a0u0442 staining or u0431u043bu043eu0442u0438u0440u043eu0432u0430u043du0438u0435 u0433u0435u043bu00a0.;14. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 1 or 2, so that u0434u043bu00a0 u043eu043fu0440u0435u0434u0435u043bu0435u043du0438u00a0 functional version of the protein are more u0438u0441u0441u043bu0435u0434u043eu0432u0430u043du0438u00a0.;15. method for 1 or 2, u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0, phase (a) involves an additional effect on protein, wild type, at least one u043fu0440u043eu0442u0435u0430u0437u043eu0439, and step (b) includes the definition of in whether, and to what extent is u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au043eu0435 fission of the wild type protein.;16. method for u043eu0442u043bu0438u0447u0430u044eu0449u0438u0439u0441u00a0 1 or 2, so that the protein of wild type and, at the discretion of a protein are u0443u0441u043bu043eu0432u0438u00a0 u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au043eu0439 reaction, but in the absence of protease (ol u043eu0442u0435u0430u0437), and then u043eu043fu0440u0435u0434u0435u043bu00a0u044eu0442 degree u043fu0440u043eu0442u0435u043eu043bu0438u0442u0438u0447u0435u0441u043au043eu0433u043e u0440u0430u0441u0449u0435u043fu043bu0435u043du0438u00a0.