Biomolecule partition motifs and uses thereof
The invention provides amino acid sequence motifs (e.g., biomolecule partition motifs) that can direct targeting of intracellular polypeptides to and through membranes, including cell surface membranes.
1. A recombinant polynucleotide that encodes a cell surface polypeptide comprising a biomolecule partition motif, wherein the biomolecule partition motif comprises the amino acid sequence of SEQ ID NO: 250 and is located at the carboxy terminus of said polypeptide.
2. The recombinant polynucleotide of claim 1 , wherein the polynucleotide is a DNA molecule.
3. The recombinant polynucleotide of claim 1 , wherein the polynucleotide is an RNA molecule.
4. A recombinant polynucleotide comprising an RNA molecule that is complementary to the RNA molecule of claim 3 .
5. The recombinant polynucleotide of claim 1 , wherein the biomolecule partition motif is SEQ ID NO: 1 (RKRSWTY).
6. An expression vector comprising a recombinant polynucleotide that encodes a cell surface polypeptide comprising a biomolecule partition motif, wherein the biomolecule partition motif comprises the amino acid sequence of SEQ ID NO: 250 and is located at the carboxy terminus of said polypeptide.
7. The expression vector of claim 6 , wherein the recombinant polynucleotide is operatively linked to an enhancer and/or promoter.
8. A transformed cell comprising the recombinant polynucleotide of claim 1 .
9. A process for preparing a transformed cell, the process comprising the steps of:
a) transfecting, electroporating or transforming the cell with a recombinant polynucleotide that encodes a cell surface polypeptide comprising a biomolecule partition motif, wherein the biomolecule partition motif comprises the amino acid sequence of SEQ ID NO: 250 and is located at the carboxy terminus of said polypeptide; and
b) maintaining the cell of step a) under conditions sufficient for expression of the polypeptide of step a).
10. A transformed cell comprising a recombinant cell surface polypeptide, wherein the recombinant cell surface polypeptide comprises a biomolecule partition motif, wherein the biomolecule partition motif comprises the amino acid sequence of SEQ ID NO: 250 and is located at the carboxy terminus of said polypeptide, and wherein the biomolecule partition motif is not present in the native polypeptide.
11. A method of obtaining a polypeptide from the surface of a cell, the method comprising:
expressing a polypeptide having at least one biomolecule partition motif in a cell under conditions sufficient to provide the polypeptide to the surface of the cell, wherein the biomolecule partition motif comprises the amino acid sequence of SEQ ID NO: 250 and is located at the carboxy terminus of said polypeptide; and
isolating the polypeptide,
thereby obtaining the polypeptide from the surface of the cell.
12. The method of claim 11 , wherein the biomolecule partition motif is SEQ ID NO: 1 (RKRSWTY).