Efficient uniform isotope labeling of Abl kinase expressed in Baculovirus-infected insect cells

被引:43
|
作者
Strauss, A [1 ]
Bitsch, F [1 ]
Fendrich, G [1 ]
Graff, P [1 ]
Knecht, R [1 ]
Meyhack, B [1 ]
Jahnke, W [1 ]
机构
[1] Novartis Inst Biomed Res, CH-4002 Basel, Switzerland
关键词
Abl kinase; Baculovirus; HSQC; NMR; STI571; uniform isotope labeling;
D O I
10.1007/s10858-005-2451-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
This report shows for the. rst time the e. cient uniform isotope labeling of a recombinant protein expressed using Baculovirus-infected insect cells. The recent availability of suitable media for N-15- and C-13/N-15-labeling in insect cells, the high expression of Abl kinase in these labeling media and a suitable labeling protocol made it possible to obtain a H-1-N-15-HSQC spectrum for the catalytic domain of Abl kinase of good quality and with label incorporation rates > 90%. The presented isotope labeling method should be applicable also to further proteins where successful expression is restricted to the Baculovirus expression system.
引用
收藏
页码:343 / 349
页数:7
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