Patent ID: 8748182

Claim:
A method of producing a mutant GPCR with increased stability in a particular conformation under denaturing conditions relative to its parent GPCR in the same particular conformation under denaturing conditions, the method comprising: (a) identifying in the amino acid sequence of one or more mutants of a first parent GPCR with increased stability in a particular conformation under denaturing conditions relative to the first parent GPCR in the same particular conformation under denaturing conditions, the position or positions at which the one or more mutants have at least one different amino acid residue compared to the first parent GPCR, and (b) making one or more mutations in the amino acid sequence that defines a second GPCR at the corresponding position or positions, to provide one or more mutants of a second parent GPCR with increased stability in a particular conformation under denaturing conditions relative to the second parent GPCR in the same particular conformation under denaturing conditions, or making one or more mutations in the amino acid sequence that defines a second GPCR within a window or windows of i plus or minus 5 residues where i is the corresponding position or positions, to provide one or more mutants of a second parent GPCR with increased stability in a particular conformation under denaturing conditions relative to the second parent GPCR in the same particular conformation under denaturing conditions, or making one or more mutations in the amino acid sequence that defines a second GPCR within a distance of 12 Å from the Cα atom of, or within a distance of 8 Å from any atom of, the amino acid residue i, where i is the corresponding position or positions, to provide one or more mutants of a second parent GPCR with increased stability in a particular conformation under denaturing conditions relative to the second parent GPCR in the same particular conformation under denaturing conditions.