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Magnetic resonance study of glycophorin A-containing13C-enriched methionines

Authors
Journal
FEBS Letters
0014-5793
Publisher
Wiley Blackwell (John Wiley & Sons)
Publication Date
Volume
143
Issue
2
Identifiers
DOI: 10.1016/0014-5793(82)80127-0
Keywords
  • 13C-Nmr
  • Glycophorin A
  • 13C-Enriched Methionines
  • [S-[Methyl-13C]Methionine] Glycophorin A
Disciplines
  • Biology
  • Chemistry

Abstract

Abstract Methionine-81 and/or -8 of the transmembrane sialoglycoprotein, glycophorin A, have been specifically alkylated with 13CH 3I 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 Me 4Si. 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 Me 4Si whereas the ε-carbon of Met-81 exhibited an unusual chemical shift of 2.0 ppm downfield from Me 4Si. The spin-lattice relaxation time of both resonances was found to be ∼0.3 s.

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