Receptor mutagenesis strategies for examination of structure-function relationships.
This chapter describes three different strategies of receptor mutagenesis with their advantages, disadvantages, and limitations. Oligonucleotide-directed mutagenesis using either the Altered Sites II in vitro mutagenesis system or the GeneTailor site-directed mutagenesis system can generate base substitutions/deletions/insertions that yield single/multiple amino acid substitutions/deletions/insertions and/or N- or C-terminal truncations in GPCRs. Polymerase chain reaction-based mutagenesis strategies allow substitutions/deletions/insertions of larger domains within GPCRs, creating truncated receptors or receptor chimeras. In addition, some guidelines are given and examples are provided to facilitate design and interpretation of mutational experiments.
Blomenröhr
Marion
M
Physiological Chemistry, Utrecht University, The Netherlands.
Vischer
Henry F
HF
Bogerd
Jan
J
D000818:Animals; D006801:Humans; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction / Q000379:methods; D043562:Receptors, G-Protein-Coupled / Q000235:genetics* / Q000378:metabolism*; D013329:Structure-Activity Relationship
D043562:Receptors, G-Protein-Coupled