Lipoparticles comprising ion channels, methods of making and using the same
The present invention relates to the use of lipoparticles, virus-like particles, and viruses. The present invention also relates to testing ion channel function and modulators of ion channels.
1. A method for identifying a test compound that can modulate ion conductance comprising:
a) contacting an isolated virus particle purified away from a cell, the particle comprising an external lipid bilayer, a viral core protein, and a selected ion conductor with a membrane potential sensitive dye and a test compound; and
b) detecting a change in membrane potential with the membrane potential sensitive dye,
wherein a modulation in ion conductance in the presence of the test compound indicates that the test compound modulates ion conductance.
2. The method of claim 1 , wherein the selected ion conductor is an ion channel polypeptide or an ionophore.
3. The method of claim 1 , further comprising activating the selected ion conductor prior to contacting the particle with the test compound.
4. The method of claim 3 , wherein the activating comprises contacting the isolated virus particle with a ligand that binds to the selected ion conductor and activates the selected ion conductor.
5. The method of claim 3 , wherein the activating comprises contacting the isolated virus particle with a salt concentration effective to activate the selected ion conductor.
6. The method of claim 1 , wherein the detecting a change in membrane potential comprises detecting a change in fluorescence.
7. The method of claim 1 , wherein the membrane potential sensitive dye is di-4-ANEPPS, di-8-ANEPPS, rhodamine 421, oxonol VI, JC-1, DiSC3(5), CC2-DMPE, DiSBAC2(3), and DiSBAC4(3), DiSBAC(1)3, FMP-Blue, FMP-Red, VABSC-1, HLB 021-152, HLB 021-155, HLB 007-054, HLB 021-149, HLB 004-111, HLB 007-052, HLB 028-008, HLB 004-078, HLB 004-183, and combinations thereof.
8. The method of claim 2 , wherein the ion channel polypeptide is hERG, SCN5a, KCNQ1+minK, hERG+MiRP1, M2, 5HT3a, Shaker, KCNQ2+KCNQ3 and KCNQ1, and combinations thereof.
9. The method of claim 1 , wherein the viral core protein is an influenza core protein, HIV core protein, SIV core protein, MLV core protein, EIAV core protein, RSV core protein, VSV core protein, ALV core protein, or baculovirus core protein.
10. The method of claim 2 , wherein the ion channel polypeptide is a viral ion channel polypeptide or a non-viral ion channel polypeptide.
11. The method of claim 10 , wherein the non-viral ion channel polypeptide is a eukaryotic ion channel polypeptide.
12. The method of claim 1 , wherein the modulation is inhibition or activation.
13. The method of claim 1 further comprising comparing the test compound's ability to modulate an ion conductor's activity to a compound known to modulate an ion channel conductor's activity.
14. The method of claim 13 , wherein the compound known to modulate an ion channel conductor's activity is amantadine or rimantadine.
15. The method of claim 1 , wherein the isolated virus particle consists of one selected ion conductor.
16. The method of claim 1 further comprising measuring the change in membrane potential.
17. The method of claim 2 , wherein the ionophore is a protonophore.
18. The method of claim 2 , wherein the ionophore is valinomycin, SQiRP, FCCP, or ionomycin.
19. The method of claim 2 , wherein the isolated virus particle comprises the ion channel polypeptide and the ionophore.
20. The method of claim 1 , wherein the virus particle is a virus-like particle.
21. The method of claim 2 , wherein the virus particle is a virus-like particle.