Inhibition of membrane fusion proteins
View Patent ↗Methods of inhibiting viral infection of a eukaryotic cell by a target virus having a class II virus fusion protein are provided. Also provided are methods of screening a test compound for the ability to inhibit infection by a virus having a class II viral fusion protein. Additionally provided herewith are aqueous-soluble proteins comprising a portion of a class II viral fusion protein comprising a Domain III of the viral fusion protein.
1. A method of screening a test compound for the ability to inhibit infection by a virus having a class II viral fusion protein, the method comprising:
(a) combining the test compound with
(i) an aqueous-soluble protein comprising a domain having the sequence set forth in SEQ ID NO:1, and
(ii) a core homotrimer of the class II viral fusion protein; and
(b) quantitating binding of the aqueous-soluble protein to the core homotrimer,
wherein reduced binding of the aqueous-soluble protein to the core homotrimer in the presence of the test compound as compared to binding of the aqueous-soluble protein to the core homotrimer in the absence of the test compound indicates that the test compound inhibits infection by the virus having the class II viral fusion protein.
2. The method of claim 1 , wherein the aqueous-soluble protein further comprises at least a portion of a stem region of an Alphavirus fusion protein, wherein the stem region comprises SEQ ID NO:2.
3. The method of claim 1 , wherein the aqueous-soluble protein further comprises a DI/DIII linker region from an Alphavirus class II fusion protein, wherein the Alphavirus linker region has SEQ ID NO: 11.
4. The method of claim 1 , wherein the aqueous-soluble protein further comprises an oligohistidine moiety.
5. The method of claim 1 , wherein the aqueous-soluble protein is labeled with a detectable label.
6. The method of claim 5 , wherein the detectable label is a fluorescent molecule, a radioactive atom, an enzyme, or an antigen not naturally occurring in the aqueous-soluble protein.
7. The method of claim 5 , wherein the detectable label is a fluorescent molecule.
8. The method of claim 7 , wherein quantitating binding of the aqueous-soluble protein and the core homotrimer in step b) is performed by fluorescence polarization.
9. The method of claim 7 , wherein quantitating binding of the aqueous-soluble protein and the core homotrimer in step b) is performed by fluorescence resonance energy transfer.
10. The method of claim 1 , wherein the core homotrimer does not comprise a Domain III of an Alphavirus fusion protein.
11. The method of claim 1 , wherein the core homotrimer is immobilized on a solid matrix.
12. The method of claim 1 , wherein the aqueous-soluble protein is immobilized on a solid matrix.