Strauss André, Bitsch Francis, Cutting Brian, Fendrich Gabriele, Graff Patrick, Liebetanz Janis, Zurini Mauro, Jahnke Wolfgang
Novartis Pharma AG, Central Technologies, Oncology Research, CH-4002 Basel, Switzerland.
J Biomol NMR. 2003 Aug;26(4):367-72. doi: 10.1023/a:1024013111478.
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 (15)N-phenylalanine, (15)N-glycine, (15)N-tyrosine or (15)N-valine. For the essential amino acids phenylalanine, tyrosine and valine high (15)N-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.
本文描述了在杆状病毒感染的昆虫细胞中成功进行蛋白质氨基酸类型选择性同位素标记的培养条件。该方法应用于用（15）N-苯丙氨酸、（15）N-甘氨酸、（15）N-酪氨酸或（15）N-缬氨酸对c-Abl激酶的催化结构域进行选择性标记。对于必需氨基酸苯丙氨酸、酪氨酸和缬氨酸，观察到（15）N标记掺入率> /= 90%，并且在HSQC谱中出现了大约预期数量的共振峰，而非必需氨基酸甘氨酸的情况并非如此。该方法应适用于其他难以进行NMR分析的重组蛋白的氨基酸类型选择性同位素标记。
