Myosin light chain kinase binding to plastic |
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Authors: | R B Pearson C House B E Kemp |
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Affiliation: | Department of Chemistry, Clemson University, Clemson, SC 29631, USA |
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Abstract: | Methionine-81 and/or -8 of the transmembrane sialoglycoprotein, glycophorin A, have been specifically alkylated with 13CH3I to produce the sulfonium ion derivatives [S-[13C]methylmethionine-8]glycophorin A and [S-[13C]methylmethionine-8 and -81]glycophorin A. 13C NMR spectra of these species show that the resonances of the methyl groups of the modified glycophorins occur at 26.1 ppm downfield from Me4Si. A spin-lattice relaxation time of 0.4 was observed for the 13C-enriched methyl resonances of the sulfonium ion derivatives of Met-8 and -81, which corresponds to an effective correlation time of < 2× 10?10 s. Demethylation of the 2 glycophorin A sulfonium ion species with 2-mercaptoethanol produces native glycophorin A which now has the ε-carbon of the methionine residue(s) 45% isotopically enriched. The ε-carbon of Met-8 was found to occur at 15.7 ppm downfield from Me4Si whereas the ε-carbon of Met-81 exhibited an unusual chemical shift of 2.0 ppm downfield from Me4Si. The spin-lattice relaxation time of both resonances was found to be ~0.3 s. |
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Keywords: | Glycophorin A α-D-NeuAc carbon-13 nuclear magnetic resonance spectroscopy PRFT partially relaxed Fourier transform SDS sodium dodecylsulfate LIS lithium diiodosalicylate |
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