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. Author manuscript; available in PMC: 2010 Nov 24.
Published in final edited form as: Biochemistry. 2009 Nov 24;48(46):10976–10987. doi: 10.1021/bi901291c

Figure 1.

Figure 1

A. The two-dimensional topology of Saccharomyces cerevisiae Ste2p. The cartoon indicates the location of the genetically engineered Factor Xa digestion site in EL2 and the epitope tags in the C-terminus. The two endogenous Cys residues (C59 and C252) were mutated to Ser to generate FT-HT-Xa which was the parental GPCR used to generate the TM1 and TM7 cysteine mutations in positions 64 to 69 in TM1 and 278 to 296 in TM7 as indicated in the boxes. B. Dose-response analysis of growth arrest assay. The zone of growth inhibition of strains carrying the indicated receptors was measured at various concentrations of α-factor. FT-HT-Xa is the Cys-less receptor containing C-terminal FLAG and His epitope tags and a tandem Factor Xa digestion site in EL2. FT-HT is the Cys-less receptor without the Xa digestion site containing the FLAG and His epitopes, and Native Ste2p is the wild-type receptor that has no epitope tags and has two Cys residues (C59 and C252).