Cell-permeable, cell-compatible, and cleavable linkers for covalent tethering of functional elements
Provided herein are cell-permeable, cell-compatible, and chemoselectively-cleavable linkers for tethering (e.g., covalently) functional elements (e.g., a cellular interaction element and a capture element), and methods of use (e.g., intracellular capture and extracellular release of cellular targets) therewith.
1. A dual-function, cell-permeable, cell-compatible, chemoselectively-cleavable composition, comprising:
(a) a cellular interaction element, wherein the cellular interaction element is capable of non-covalently binding to a cellular target protein;
(b) a capture element comprising a haloalkane group; and
(c) a linker covalently tethering the cellular interaction element to the capture element, wherein said linker comprises a chemoselectively cleavable moiety selected from the group consisting of an allyl-heteroatom group, a propargyl-heteroatom group.
2. The composition of claim 1 , wherein the cellular interaction element is a small molecule or peptide.
3. The composition of claim 1 , wherein the capture element is a covalent substrate for an enzyme.
4. The composition of claim 1 , wherein the capture element is an affinity element.
5. The composition of claim 1 , wherein the chemoselectively cleavable moiety comprises an allyl-heteroatom group selected from the group consisting of: allyl ether, allyl amine, allyl ester, allyl amide, allyl urea, allyl carbonate, and allyl carbamate.
6. The composition of claim 5 , wherein the chemoselectively-cleavable moiety comprises an allyl carbamate group.
7. The composition of claim 1 , wherein the chemoselectively-cleavable moiety comprises a propargyl-heteroatom group selected from the group consisting of: propargyl ether, propargyl amine, propargyl ester, propargyl amide, propargyl urea, propargyl carbonate, and propargyl carbamate.
8. A method comprising:
(a) administering the dual-function, cell-permeable, cell compatible, chemoselectively-cleavable composition of claim 1 to a cell;
(b) allowing binding of the chemoselectively-cleavable composition via the cellular interaction element with a corresponding intracellular cellular target to provide chemoselectively-cleavable composition bound intracellular target;
(c) lysing said cell to produce a cell lysate; and
(d) providing surface immobilized intracellular target by contacting the cell lysate with a surface immobilized capture agent specific for the capture element of the chemoselectively-cleavable composition to immobilize the chemoselectively-cleavable composition bound intracelluar target on the surface.
9. The method of claim 8 , further comprising:
(e) contacting the surface immobilized intracellular target with a chemoselective agent capable of cleaving the chemoselectivly-cleavable linker, thereby releasing the cellular interaction element and the bound intracellular target from the surface.
10. A system comprising:
(a) the dual-function, cell-permeable, cell-compatible, chemoselectively-cleavable composition of claim 1 ; and
(b) one or more of:
(i) a cell,
(ii) a cellular target,
(iii) a surface-displayed capture agent, and
(iv) a chemoselective agent.