The invention relates to polymeric compounds of formula I which allow the solubilization and/or stabilization of membrane proteins, it relates in particular to the complexes they form with membrane proteins and the method of solubilizing and/or stabilizing membrane proteins comprising the use of these compounds.
1 . A water-soluble membrane protein-amphiphilic vinyl polymer complex comprising the amphiphilic vinyl polymer of formula I:
wherein:
M + is an alkali metal,
R 1 , R 4 , R 5 , R 8 are each independently a hydrogen atom or a methyl radical,
R 2 , R 6 , R 9 are each independently a group selected from a hydrogen atom, linear (C1-C8) alkyls, and branched (C3-C8) alkyls,
R 10 is a group selected from linear (C1-C5) alkyls and branched (C3-C5) alkyls,
R 3 and R 7 are each independently a group selected from the group consisting of:
hydrogen, with X then being a single bond,
linear or branched alkyls of at least 6 carbon atoms, linear or branched alkenyls of at least 6 carbon atoms, linear or branched alkynyls of at least 6 carbon atoms, with X then being a single bond,
(C3-C10) cycloalkyls or (C3-C10) (hetero) cycloalkyls, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls, with X then being selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (3-C8) alkenylenes, linear (C2-C8) alkynylenes and branched (C3-C8) alkynylenes,
(C3-C10) cycloalkenyls or (C3-C10) (hetero) cycloalkenyls, poly or monounsaturated, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, linear or branched (C2-C8) alkynyls, with X then being selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (C2-C8) alkenylenes, linear (C2-C8) alkynylenes and branched (C2-C8) alkynylenes,
poly or monounsaturated polycycles, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls, with X then being selected from a single bond, linear (C2-C8) alkylenes or branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (C2-C8) alkenylenes, linear (C2-C8) alkynylenes and branched (C3-C8) alkynylenes, and
phenyl, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls, with X then being selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (C3-C8) alkenylenes, linear (C2-C8) alkynylenes, and branched (C3-C8) alkynylenes,
with w, x, y, z corresponding to the respective percentages of the units,
with x being between 20 and 90%,
with y being between 0 and 80%,
with w being between 0 and 80%, and
with z being between 0 and 60%, provided that w+x+y+z=100%, and that y or w is at least 10%,
provided that when one of the groups R 3 or R 7 is a linear or branched alkyl of at least 6 carbon atoms, a linear or branched alkenyl of at least 6 carbon atoms, a linear or branched alkynyl of at least 6 carbon atoms, or a hydrogen atom, with X being a single bond, then w and y are different from 0 and the other group R 3 or R 7 is selected from the group consisting of:
C3-C10) cycloalkyls or (C3-C10) (hetero) cycloalkyls, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls, with X then being selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (C3-C8) alkenylenes, linear (C2-C8) alkenylenes and branched (C3-C8) alkynylenes,
(C3-C10) cycloalkenyls or (C3-C10) (hetero) cycloalkenyls, poly or monounsaturated, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls, with X then being selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes or branched (C3-C8) alkenylenes, linear (C2-C8) alkynylenes, and branched (C3-C8) alkynylenes,
poly or monounsaturated polycycles, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls, with X then being selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (C3-C8) alkenylenes, linear (C2-C8) alkynylenes, and branched (C3-C8) alkynylenes, and
phenyl, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls, with X then being selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (C3-C8) alkenylenes, linear (C2-C8) alkynylenes, and branched (C3-C8) alkynylenes.
2 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein in the amphiphilic vinyl polymer of formula I, M + is Na + .
3 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein in the amphiphilic vinyl polymer of formula I, at least one of w or y is greater than or equal to 25% and less than or equal to 80%.
4 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein in the amphiphilic vinyl polymer of formula I, R 10 is an isopropyl group.
5 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein in the amphiphilic vinyl polymer of formula I, x is at least 20% and less than or equal to 75%.
6 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein in the amphiphilic vinyl polymer of formula I, at least one of w or y is between 25% and 50%, and z is between 0% and 40%.
7 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein in the amphiphilic vinyl polymer of formula I:
at least one of R 3 or R 7 is (C 6 -C 8 ) cycloalkyls, or
at least one of R 3 or R 7 is phenyl,
with X being selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (C3-C8) alkenylenes, linear (C2-C8) alkynylenes, and branched (C3-C8) alkynylenes.
8 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein in the amphiphilic vinyl polymer of formula I:
each of R 1 , R 2 , R 4 , R 5 , R 6 , R 8 , and R 9 , is a hydrogen atom,
at least one of R 3 or R 7 is (C 6 -C 8 ) cycloalkyl, and X is a single bond,
R 10 is an isopropyl group,
x is equal to 35% or 50%,
y is greater than or equal to 25%,
w is equal to 0%, or 25%, and
z is equal to 0%, 15% or 40%, provided that w+x+y+z=100%.
9 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein the amphiphilic vinyl polymer is selected from:
10 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein the amphiphilic vinyl polymer of formula I is functionalized by grafting an affinity tag, a fluorescent probe, or an immunostimulatory molecule.
11 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , further comprising lipid compounds.
12 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , wherein M+ is selected from Li + , Na + , K + .
13 . The water-soluble membrane protein-amphiphilic vinyl polymer complex according to claim 1 , said phenyl being para substituted with a methyl or ethyl group.
14 . An aqueous solution comprising the complex according to claim 1 , comprising a mixture of membrane proteins, including at least one stabilized membrane protein.
15 . A process of preparing the solution according to claim 14 , comprising a solubilization and/or stabilization step in which a protein fraction from a biological or synthetic membrane containing a said membrane protein, or said mixture of membrane proteins, is brought into the presence of said vinyl polymer of formula I.
16 . The process of preparing the solution according to claim 15 , wherein said membrane protein, or said mixture of membrane proteins, is/are selected from among: one or more recombinant proteins expressed in prokaryotic or eukaryotic cells, one or more native proteins expressed in specialized membranes with high protein density, and a mixture thereof.
17 . The process of preparing the solution according to claim 16 , comprising first solubilizing said membrane protein, or said mixture of membrane proteins, by bringing the membrane protein fraction or the mixture of membrane proteins together with a detergent medium.
18 . The process of preparing the solution according to claim 16 , comprising first fusing the protein fraction containing a said membrane protein, or said mixture of membrane proteins, with vesicles of phospholipids or of a mixture of phospholipids.
19 . The water process of preparing the solution according to claim 18 , said vesicles comprising 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC).
20 . The process of preparing the solution according to claim 16 , wherein the protein fraction containing said membrane protein is not a solution of membrane protein(s) in a detergent medium in micellar form.
21 . A polymer of formula I:
wherein:
M + is an alkali metal,
with w, x, y, z corresponding to the respective percentages of the units,
with x being between 20 and 90%,
with y being between 10 and 80%,
with w being 0%, and
with z being between 0 and 60%, provided that w+x+y+z=100%%,
R 4 , R 5 , R 8 are each independently a hydrogen atom or a methyl radical,
R 6 , R 9 are each independently a group selected from a hydrogen atom, linear (C1-C8) alkyls, and branched (C3-C8) alkyls,
R 10 is a group selected from linear (C1-C5) alkyls and branched (C3-C5) alkyls,
R7 is a group selected from the group consisting of:
(C3-C10) cycloalkyls or (C3-C10) (hetero) cycloalkyls, unsubstituted or substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls, and
phenyl, substituted by one or more radicals selected from linear, cyclic or branched (C1-C8) alkyls, linear or branched (C2-C8) alkenyls, and linear or branched (C2-C8) alkynyls,
and wherein X is selected from a single bond, linear (C2-C8) alkylenes, branched (C3-C8) alkylenes, linear (C2-C8) alkenylenes, branched (C3-C8) alkenylenes, linear (C2-C8) alkylenes, and branched (C3-C8) alkynylenes.