Speaker: Dr. Gary J. Pielak
The University of North Carolina at Chapel Hill, USA

HOST

Miquel Pons, IRB Barcelona /UB
Friday, 2 July 2010, 12:00h Aula Fèlix Serratosa

ABSTRACT

Although over expression and 15N enrichment facilitate the observation of resonances from disordered proteins in Escherichia coli, 15N enrichment alone is insufficient for detecting most globular proteins. I will explain this dichotomy and describe how we overcame the problem while extending the capability of in-cell NMR by using 19F-labeled proteins. Resonances from small (〜10 kDa) globular proteins containing the amino acid analogue 3-fluoro-tyrosine can be observed in cells, but for larger proteins the 19F resonances are broadened beyond detection. Incorporating the amino acid analogue trifluoromethyl-L-phenylalanine allows larger proteins (up to 100 kDa) to be observed in cells.

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