Amino-acid-type selective isotope labeling of proteins expressed in Baculovirus-infected insect cells useful for NMR studies |
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Authors: | Strauss André Bitsch Francis Cutting Brian Fendrich Gabriele Graff Patrick Liebetanz Janis Zurini Mauro Jahnke Wolfgang |
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Affiliation: | (1) Novartis Pharma AG, Central Technologies, Oncology Research, CH-4002 Basel, Switzerland |
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Abstract: | Culture conditions for successful amino–acid-type selective isotope labeling of proteins expressed in Baculovirus-infected insect cells are described. The method was applied to the selective labeling of the catalytic domain of c-Abl kinase with 15N-phenylalanine, 15N-glycine, 15N-tyrosine or 15N-valine. For the essential amino acids phenylalanine, tyrosine and valine high 15N-label incorporation rates of 90% and approximately the expected number of resonances in the HSQC spectra were observed, which was not the case for the non-essential amino acid glycine. The method should be applicable to amino-acid-type selective isotope labeling of other recombinant proteins which have not been amenable to NMR analysis. |
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Keywords: | Abl kinase Baculovirus HSQC NMR selective labeling STI-571 |
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