Document Detail


Biosynthetically directed (2)H labelling for stereospecific resonance assignments of glycine methylene groups.
MedLine Citation:
PMID:  23192292     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Stereospecific resonance assignments of the α-protons of glycine are often difficult to obtain by measurements of scalar coupling constants or nuclear Overhauser effects. Here we show that these stereospecific resonance assignments can readily be obtained by cell-free protein synthesis in D(2)O, as the serine hydroxymethyltransferase, that is naturally present in E. coli cell extracts, selectively replaces the pro-2S proton of glycine by a deuterium. To encourage the conversion by serine hydroxymethyltransferase, we performed the cell-free reaction without the addition of any glycine, exploiting the capability of the enzyme to convert serine to glycine with the help of tetrahydrofolate. (13)C-HSQC spectra of ubiquitin produced with (13)C/(15)N-serine showed that about a quarter of the glycine residues derived from serine were stereospecifically deuterated. Pulse sequences are presented that select the signals from the stereospecifically deuterated glycine residues.
Authors:
Karin V Loscha; Gottfried Otting
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-11-29
Journal Detail:
Title:  Journal of biomolecular NMR     Volume:  -     ISSN:  1573-5001     ISO Abbreviation:  J. Biomol. NMR     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-11-29     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9110829     Medline TA:  J Biomol NMR     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Research School of Chemistry, Australian National University, Canberra, ACT, 0200, Australia.
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