Glycan-Tethered Stabiligases
AssigneeThe Regents of the University of California
InventorsJames A. Wells, Kaitlin Schaefer
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Abstract
Provided herein are novel stabiligases for use in cell membrane proteome analysis. The subject stabiligases are capable of attaching to glycans found on the surface of cell membranes to form glycan-tethered (GT) stabiligases. Such glycan-tethered stabiligases are capable of robustly and selectively attaching label probes to cell surface proteins of intact cells. The subject novel stabiligases described herein advantageously allow for the identification and profiling of cell membrane proteins that have undergone an extracellular N-terminal proteolytic event.
Claims20
B1-B3. (canceled)
C1-C7. (canceled)
A cell comprising a stabiligase tethered to extracellular glycans on intact human cells.
A method of making a cell comprising a membrane-tethered stabiligase, comprising oxidative coupling a stabiligase to a cell-membrane protein on the surface of the cell.
A stabiligase comprising an N-terminal an α-aminooxy- or α-hydrazido-group.
A method of identifying a population of cell membrane proteins on a cell, the method comprising:
- a) providing a cell comprising a membrane-tethered stabiligase and a population of cell membrane proteins;
- b) contacting the cell with a label probe comprising an ester substrate and a detectable label under conditions wherein the membrane-tethered stabiligase attaches the label probe to the population of cell membrane proteins to form a population labelled cell membrane proteins;
- c) isolating the labelled cell membrane proteins;
- and
- d) analyzing the isolated labelled cell membrane proteins;
- therein identifying the population of cell membrane proteins on the cell.
Dependencies are inferred from the claim text.