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- W2055722719 abstract "To conduct biophysical analyses on large domains of GPCRs, multimilligram quantities of highly homogeneous proteins are necessary. This communication discusses the biosynthesis of four transmembrane and five transmembrane-containing fragments of Ste2p, a GPCR recognizing the Saccharomyces cerevisiae tridecapeptide pheromone α-factor. The target fragments contained the predicted four N-terminal Ste2p[G31-A198] (4TMN), four C-terminal Ste2p[T155-L340] (4TMC), or five C-terminal Ste2p[I120-L340] (5TMC) transmembrane segments of Ste2p. 4TMN was expressed as a fusion protein using a modified pMMHa vector in L-arabinose-induced Escherichia coli BL21-AI, and cleaved with cyanogen bromide. 4TMC and 5TMC were obtained by direct expression using a pET21a vector in IPTG-induced E. coli BL21(DE3) cells. 4TMC and 5TMC were biosynthesized on a preparative scale, isolated in multimilligram amounts, characterized by MS and investigated by biophysical methods. CD spectroscopy indicated the expected highly α-helical content for 4TMC and 5TMC in membrane mimetic environments. Tryptophan fluorescence showed that 5TMC integrated into the nonpolar region of 1-stearoyl-2-hydroxy-sn-glycero-3-phospho-(1′-rac-glycerol) micelles. HSQC-TROSY investigations revealed that [15N]-labeled 5TMC in 50% trifluoroethanol-d2/H2O/0.05%–trifluoroacetic acid was stable enough to conduct long multidimensional NMR measurements. The entire Ste2p GPCR was not readily reconstituted from the first two and last five or first three and last four transmembrane domains. © 2012 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 98: 485–500, 2012." @default.
- W2055722719 created "2016-06-24" @default.
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- W2055722719 date "2012-01-01" @default.
- W2055722719 modified "2023-09-26" @default.
- W2055722719 title "Large multiple transmembrane domain fragments of a G protein-coupled receptor: Biosynthesis, purification, and biophysical studies" @default.
- W2055722719 cites W1683272306 @default.
- W2055722719 cites W1753435687 @default.
- W2055722719 cites W1825353903 @default.
- W2055722719 cites W1908561802 @default.
- W2055722719 cites W1965489457 @default.
- W2055722719 cites W1967925189 @default.
- W2055722719 cites W1968533552 @default.
- W2055722719 cites W1969949441 @default.
- W2055722719 cites W1970861588 @default.
- W2055722719 cites W1971026764 @default.
- W2055722719 cites W1975085160 @default.
- W2055722719 cites W1975740938 @default.
- W2055722719 cites W1977509789 @default.
- W2055722719 cites W1978167732 @default.
- W2055722719 cites W1980823735 @default.
- W2055722719 cites W1980962873 @default.
- W2055722719 cites W1981693484 @default.
- W2055722719 cites W1997729584 @default.
- W2055722719 cites W2000565142 @default.
- W2055722719 cites W2000893487 @default.
- W2055722719 cites W2005222613 @default.
- W2055722719 cites W2006212946 @default.
- W2055722719 cites W2007089493 @default.
- W2055722719 cites W2008383753 @default.
- W2055722719 cites W2014055152 @default.
- W2055722719 cites W2014550271 @default.
- W2055722719 cites W2015533931 @default.
- W2055722719 cites W2016770664 @default.
- W2055722719 cites W2028291036 @default.
- W2055722719 cites W2029748523 @default.
- W2055722719 cites W2036931150 @default.
- W2055722719 cites W2037198168 @default.
- W2055722719 cites W2037260827 @default.
- W2055722719 cites W2037287070 @default.
- W2055722719 cites W2037598310 @default.
- W2055722719 cites W2039818756 @default.
- W2055722719 cites W2041566506 @default.
- W2055722719 cites W2045102779 @default.
- W2055722719 cites W2045884212 @default.
- W2055722719 cites W2047144511 @default.
- W2055722719 cites W2047605004 @default.
- W2055722719 cites W2052029379 @default.
- W2055722719 cites W2053188903 @default.
- W2055722719 cites W2056403651 @default.
- W2055722719 cites W2057437570 @default.
- W2055722719 cites W2062973238 @default.
- W2055722719 cites W2063233717 @default.
- W2055722719 cites W2068227670 @default.
- W2055722719 cites W2069602336 @default.
- W2055722719 cites W2078434336 @default.
- W2055722719 cites W2088193170 @default.
- W2055722719 cites W2090643737 @default.
- W2055722719 cites W2100443187 @default.
- W2055722719 cites W2104477830 @default.
- W2055722719 cites W2104961861 @default.
- W2055722719 cites W2109939279 @default.
- W2055722719 cites W2117003397 @default.
- W2055722719 cites W2117345265 @default.
- W2055722719 cites W2130783152 @default.
- W2055722719 cites W2143061023 @default.
- W2055722719 cites W2149963584 @default.
- W2055722719 cites W2152853006 @default.
- W2055722719 cites W2155350436 @default.
- W2055722719 cites W2157558738 @default.
- W2055722719 cites W2157663874 @default.
- W2055722719 cites W2157959080 @default.
- W2055722719 cites W2160171025 @default.
- W2055722719 cites W2160259388 @default.
- W2055722719 cites W2166085305 @default.
- W2055722719 cites W2170665326 @default.
- W2055722719 cites W2170963882 @default.
- W2055722719 cites W2171501765 @default.
- W2055722719 cites W2599059709 @default.
- W2055722719 cites W4233523656 @default.
- W2055722719 cites W4238558616 @default.
- W2055722719 doi "https://doi.org/10.1002/bip.22122" @default.
- W2055722719 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3542537" @default.
- W2055722719 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/23203693" @default.
- W2055722719 hasPublicationYear "2012" @default.
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