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Suppression of isotope scrambling in cell-free protein synthesis by broadband inhibition of PLP enzymes for selective 15 N-labelling and production of perdeuterated proteins in H 2 O

Su, Xun-Cheng; Loh, Choy; Qi, Ruhu; Otting, Gottfried

Description

Selectively isotope labelled protein samples can be prepared in vivo or in vitro from selectively labeled amino acids but, in many cases, metabolic conversions between different amino acids result in isotope scrambling. The best results are obtained by cell-free protein synthesis, where metabolic enzymes are generally less active, but isotope scrambling can never be suppressed completely. We show that reduction of E. coli S30 extracts with NaBH4 presents a simple and inexpensive way to achieve...[Show more]

CollectionsANU Research Publications
Date published: 2011
Type: Journal article
URI: http://hdl.handle.net/1885/67647
Source: Journal of Biomolecular NMR
DOI: 10.1007/s10858-011-9477-5

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