Method for adjusting ras ubiquitination
AssigneeXiamen University
InventorsHongrui WANG, Xianming DENG, Taoling ZENG, Tingting JIANG
Life of this patent
Estimating the term…
Abstract
Provided is use of E3 ubiquitin ligase Nedd4-1 in preparation of a drug for inhibiting the Ras protein level. Also provided is use in preparation of a drug for preventing and/or treating tumors or a drug for prolonging the life span of organisms.
Claims10
A method of identifying a compound that regulates Ras level, the method comprising:
- a. contacting a Ras-expressing cell or an in vitro ubiquitination system with a test compound;
- b. selecting a compound that can increase or decrease the Ras level of the cell or system in step a, preferably selecting a compound that can decrease the Ras level of the cell or system in step a;
- optionally, the method further comprising:
- c. detecting whether the compound obtained in step b regulates the Ras level depending on the presence of Nedd4-1 protein or not;
- and
- d. selecting a compound that regulates the Ras level depending on the presence of the Nedd4-1 protein.
A method of regulating Ras level in a cell, the method comprising:
- a. contacting a compound that regulates ubiquitination degradation of Ras with a Ras-expressing cell or an in vitro ubiquitination system, optionally the activity of the compound that regulates ubiquitination degradation of Ras is Nedd4-1 dependent or independent;
- b. optionally, further regulating the level of Nedd4-1 in a cell, optionally regulating the level of Nedd4-1 comprising:
- a. gene editing, b. overexpression, c. RNA interference, d. a small molecule compound, and e. an antibody.
A method of regulating Ras level, the method comprising:
- a. judging the state of Ras protein;
- b. if the Ras protein is a wild-type protein or in an inactive state, regulating the Ras level by regulating the expression level of Nedd4-1;
- and
- c. if the Ras protein is a mutant protein or in an active state, regulating the Ras level by adding a compound;
- wherein
- optionally, regulating the expression level of Nedd4-1 and adding the compound can be performed simultaneously;
- optionally, the Ras level is decreased by up-regulating the level of Nedd4-1, or increased by down-regulating the level of Nedd4-1;
- optionally, the level of Ras protein is decreased by adding a compound without affecting the mRNA level of Ras.
The method of any one of the preceding claims, wherein the Ras is selected from H-Ras, K-Ras or N-Ras or a mutant thereof, preferably the mutant is selected from G12V, G12D, G13V, G13D and Q61R, more preferably the mutant is G12V.
A kit comprising one or more reagents for detecting Ras or Nedd4-1 level, optionally, further comprising an expression plasmid expressing Ras or Nedd4-1, optionally, further comprising a host cell expressing Ras, optionally, further comprising a compound promoting ubiquitination degradation of Ras, optionally, the compound being a compound obtained by using the method of any one of the preceding claims of the invention.
Dependencies are inferred from the claim text.