Tandem facial amphiphiles
The invention provides tandem facial amphiphiles for biochemical manipulations and characterization of membrane proteins, such as intrinsic membrane proteins. Members of this new family display favorable behavior with several membrane proteins. These amphiphiles can form relatively small micelles, and small changes in amphiphile chemical structures can result in large changes in their physical properties. The tandem facial amphiphiles can be used to aid the solubilization, isolation, purification, stabilization, crystallization, and/or structural determination of membrane proteins.
1. A method of solubilizing or stabilizing a membrane protein comprising contacting a membrane protein with an effective amount of a plurality of compounds, in an aqueous solution, and optionally heating the protein and the compounds, thereby forming a solubilized or stabilized aggregation of the compounds and the membrane protein, wherein the plurality of compounds comprises compounds of Formula I:
wherein
L is (C 5 -C 8 )cycloalkyl; phenyl optionally by 1-4 (C 1 -C 4 )alkyl; —C(R x ) 2 —; or —CH 2 —C(R x ) 2 —CH 2 —; where each R x is independently OH, or —CH 2 O-Sac;
each X is independently O, S, NH, triazole; or a direct bond;
each m is 0 or 1;
each R 1 is independently H or (C 1 -C 20 )alkyl; and
each Amph is independently a moiety of Formula A:
wherein
Y is CH 2 or a direct bond;
each R 2 , R 3 , and R 4 is independently H, OH, or O-Sac; and
each Sac is independently a monosaccharide, disaccharide, or trisaccharide;
wherein each of the compounds has at least 4 Sac groups.
2. The method of claim 1 wherein the aggregation of the compounds is in the form of a micelle.
3. The method of claim 1 wherein each R 1 is (C 1 -C 20 )alkyl.
4. The method of claim 1 wherein Y is a direct bond.
5. The method of claim 1 wherein at least two of R 2 , R 3 and R 4 are O-Sac and each Sac is a disaccharide.
6. The method claim 1 wherein the plurality of compounds comprises a compound of Formula III:
7. The method claim 1 wherein the plurality of compounds comprises a compound of Formula XII:
where each R 2 is independently an oxygen-linked monosaccharide, disaccharide, or trisaccharide.
8. The method claim 7 wherein the plurality of compounds comprises:
9. The method claim 1 wherein the plurality of compounds comprises a compound of Formula XIII:
10. The method claim 9 wherein the plurality of compounds comprises:
11. The method claim 1 wherein the plurality of compounds comprises a compound of Formula XIV:
where each R 2 is independently a monosaccharide, disaccharide, or trisaccharide.
12. The method claim 11 wherein the plurality of compounds comprises:
13. A method of solubilizing or stabilizing a membrane protein comprising contacting a membrane protein with an effective amount of a plurality of compounds, in an aqueous solution, and optionally heating the protein and the compounds, thereby forming a solubilized or stabilized aggregation of the compounds and the membrane protein, wherein the plurality of compounds comprises compounds of Formula I:
wherein
L is (C 5 -C 8 )cycloalkyl; phenyl optionally substituted by 1-4 (C 1 -C 4 )alkyl; —(CH 2 ) n — where n is 1, 2, or 4-10; —C(R x ) 2 —; or —CH 2 —C(R x ) 2 —CH 2 —; where each IV is independently OH, or —CH 2 O-Sac;
each X is independently O, S, NH, triazole; or a direct bond;
each m is 0;
each R 1 is independently H or (C 1 -C 20 )alkyl; and
each Amph is independently a moiety of Formula A:
wherein
Y is CH 2 or a direct bond;
each R 2 , R 3 , and R 4 is independently H, OH, or O-Sac; and
each Sac is independently a monosaccharide, disaccharide, or trisaccharide;
wherein each of the compounds has at least 4 Sac groups.
14. The method of claim 13 wherein the aggregation of the compounds is in the form of a micelle.
15. The method of claim 13 wherein Y is a direct bond.
16. The method of claim 13 wherein at least two of R 2 , R 3 and R 4 are O-Sac and each Sac is a disaccharide.
17. The method claim 13 wherein the plurality of compounds comprises a compound of Formula XIII:
18. The method claim 17 wherein the plurality of compounds comprises:
19. The method claim 13 wherein the plurality of compounds comprises a compound of Formula XIV:
where each R 2 is independently a monosaccharide, disaccharide, or trisaccharide.
20. The method claim 19 wherein the plurality of compounds comprises: