Back to results
US 2011/0031438 A1Published application
Methods and compositions for obtaining high-resolution crystals of membrane proteins
AssigneeThe Scripps Research Institute
InventorsRaymond C. Stevens, Michael A. Hanson, Vadim Cherezov, Peter Kuhn
Life of this patent
Estimating the term…
Abstract
The invention describes compositions and method useful for the crystallization of membrane proteins.
Claims47
A composition for lipidic cubic phase crystallization of a membrane protein, comprising
- a polyethylene glycol or modified polyethylene glycol;
- 0.01-1M of a salt;
- a host lipid;
- a lipid additive, wherein said lipid additive is present at 10-60% v/v ratio relative to the host lipid;
- a buffer;
- and
- 1 to 100 mg/ml of a membrane protein.
4 more elements
The composition of claim 28 or 29, further comprising a ligand selected from the group consisting of carazolol, timolol, alprenolol, and clenbutorol.
A method of generating crystals of a membrane protein comprising:
- mixing a lipid additive with a host lipid to form a lipid mixture, wherein said lipid additive is selected from the group consisting of a sterol, DOPE, DOPE-Me, DOPC, and Asolectin, and wherein said lipid additive is 5 to 20% w/w in said host lipid;
- and
- combining said lipid mixture with a membrane protein solution under conditions suitable for the formation of a lipidic cubic phase composition.
A method of screening a crystal of a GPCR present in a liquid cubic phase composition comprising:
- preparing a liquid cubic phase composition comprising a GPCR protein, a host lipid, and a lipid additive;
- exposing said composition to a first X-ray beam and determining a change in direction or intensity of said first X-ray beam;
- exposing said composition to a second beam and determining a change in direction or intensity of said second X-ray beam;
- identifying an area where said GPCR crystal is present in said composition;
- and
- exposing said identified area to at least a third X-ray beam.
3 more elements
Dependencies are inferred from the claim text.