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Mutagenesis Approaches for Elucidation of Protein Structure‐Function Relationships

Catherine D. Strader¹

¹Schering‐Plough Research Institute, Kenilworth, New Jersey

Publication Name:

Current Protocols in Neuroscience

Unit Number:

Unit 4.9

DOI:

10.1002/0471142301.ns0409s03

Online Posting Date:

May, 2001

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Abstract

Several mutagenesis strategies have been used to allow the identification of specific receptor-ligand interactions to support the docking of ligands within various receptor models. The strengths and limitations of each strategy are discussed in this overview. Large-scale mapping is described using deletion and chimeric receptors. Fine tuning with single residue replacements is also covered along with two-dimensional approaches.

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Unit Introduction
Large-Scale Mapping: Deletions and Chimeric Receptors
Fine-Tuning with Single-Residue Replacements and Two-Dimensional Approaches
Conclusion
Literature Cited

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Literature Cited

Literature Cited
	Dixon, R.A.F., Sigal, I.S., Rands, E., Register, R.B., Candelore, M.R., Blake, A.D., and Strader, C.D. 1987. Ligand binding to the -adrenergic receptor involves its rhodopsin-like core. Nature 326:73-77.
	Fong, T.M., Cascieri, M.A., Yu, H., Bansal, A., Swain, C., and Strader, C.D. 1993. Amino-aromatic interaction between histidine-197 of the human neurokinin-1 receptor and CP-96,345. Nature 362:350-353.
	Fong, T.M., Yu, H., Cascieri, M.A., Underwood, D., Swain, C.J., and Strader, C.D. 1994. The role of histidine-265 in antagonist binding to the neurokinin-1 receptor. J. Biol. Chem. 269:2728-2732.
	Gether, U., Johansen, T.E., Snider, R.M., Lowe, J.A., Nakanishi, S., and Schwartz, T.W. 1993. Different binding epitopes on the NK₁ receptor for substance P and a non-peptide antagonist. Nature 362:345-348.
	Guan, X-M., Perontka, S.J., and Kobilka, B.K. 1992. Identification of a single amino acid residue responsible for the binding of a class of -adrenergic receptor antagonists to 5-hydroxy tryptamine-1A receptors. Mol. Pharmacol. 41:695-698.
	Rucker, J., Samson, M., Doranz, B.J., Libert, F., Berson, J.F., Yi, Y., Smyth, R.J., Collman, R.G., Broder, C.C., Vassart, G., Doms, R.W. and Parmentier, M. 1996. Regions in -chemokine receptors CCR5 and CCR2b that determine HIV-1 cofactor specificity. Cell 87:437-446.
	Schertler, G.F., Villa, C., and Henderson, R. 1993. Projection structure of rhodopsin. Nature 362:770-772.
	Strader, C.D., Sigal, I.S., Candelore, M.R., Rands, E., Hill, W.S., and Dixon, R.A.F. 1988. Conserved aspartic acid residues 79 and 113 of the -adrenergic receptor have different roles in receptor function. J. Biol. Chem. 263:10267-10271.
	Strader, C.D., Candelore, M.R., Hill, W.S., Sigal, I.S., and Dixon, R.A.F. 1989. Identification of two serine residues involved in agonist activation of the -adrenergic receptor. J. Biol. Chem. 264:13572-13578.
	Strader, C.D., Gaffney, T., Sugg, E., Candelore, M., Keys, R., Patchett, A., and Dixon, R. 1991. Allele-specific activation of genetically engineered receptors. J. Biol. Chem. 256:5-8.
	Strader, C.D., Fong, T.M., Tota, M.R. Underwood, D., and Dixon, R.A.F. 1994. Structure and function of G protein–coupled receptors. Annu. Rev. Biochem. 63:101-132.