Efficient uniform isotope labeling of Abl kinase expressed in Baculovirus-infected insect cells.
This report shows for the first time the efficient uniform isotope labeling of a recombinant protein expressed using Baculovirus-infected insect cells. The recent availability of suitable media for (15)N- and (13)C/(15)N-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 (1)H-(15)N-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.
Strauss
André
A
Novartis Institutes for BioMedical Research, Basel, Switzerland. andre.strauss@novartis.com
Bitsch
Francis
F
Fendrich
Gabriele
G
Graff
Patrick
P
Knecht
René
R
Meyhack
Bernd
B
Jahnke
Wolfgang
W
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism*; D016466:CHO Cells; D002247:Carbon Isotopes; D020134:Catalytic Domain; D002460:Cell Line; D002470:Cell Survival; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007313:Insecta; D009682:Magnetic Resonance Spectroscopy; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D016315:Proto-Oncogene Proteins c-abl / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins; D016315:Proto-Oncogene Proteins c-abl