Abstract
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 15N- and 13C/15N-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 1H–15N-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.
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S.J. Archer A. Bax A.B. Roberts M.B. Sporn Y. Ogawa K.A. Piez J.A. Weatherbee M.L. Tsang R. Lucas B. Zheng J. Wenker D.A. Torchia (1993) Biochemistry 32 1152–1163 Occurrence Handle10.1021/bi00055a021 Occurrence Handle1:CAS:528:DyaK3sXlt1KjtA%3D%3D Occurrence Handle8424942
A. Bax (1994) Curr. Opin. Struct. Biol. 4 738–744 Occurrence Handle10.1016/S0959-440X(94)90173-2 Occurrence Handle1:CAS:528:DyaK2cXmvFKhsLg%3D
G. Bold J. Frei P. Furet P.W. Manley J. Bruggen R. Cozens S. Ferrari F. Hofmann G. Martiny-Baron J. Mestan T. Meyer J.M. Wood (2002) Drug of the Future 27 43–55 Occurrence Handle1:CAS:528:DC%2BD38XjtVCgtbs%3D
M. Brüggert T. Rehm S. Shanker J. Georgescu T.A. Holak (2003) J. Biomol. NMR 25 335–348 Occurrence Handle12766395
B. Cutting A. Strauss G. Fendrich P.W. Manley W. Jahnke (2004) J. Biomol. NMR 30 205–210 Occurrence Handle1:CAS:528:DC%2BD2MXnvVKkug%3D%3D Occurrence Handle15702527
D. Fabbro S. Ruetz E. Buchdunger S.W. Cowan-Jacob G. Fendrich J. Liebetanz J. Mestan T. O’Reilly P. Traxler B. Chaudhuri H. Fretz J. Zimmermann T. Meyer G. Caravatti P. Furet P.W. Manley (2002) Pharmacol. Therapeut. 93 79–98 Occurrence Handle1:CAS:528:DC%2BD38XksVShtbw%3D
A.P. Hansen A.M. Petros A.P. Mazar T.M. Pederson A. Rueter S.W. Fesik (1992) Biochemistry 51 12713–12718
D.C. Muchmore L.P. McIntosh C.B. Russell D.E. Anderson F.W. Dahlquist (1989) Method Enzymol. 177 44–73 Occurrence Handle1:CAS:528:DyaK3cXktVarur4%3D
B. Nagar W.G. Bornmann P. Pellicena T. Schindler D.R. Veach W.T. Miller B. Clarkson J. Kuriyan (2002) Cancer Res. 62 4236–4243 Occurrence Handle1:CAS:528:DC%2BD38XlvFeltLY%3D Occurrence Handle12154025
D.R. O’Reilly L.K. Miller V.A. Luckow (1994) Baculovirus Expression Vectors, Oxford University Press New York, NY
A. Strauss F. Bitsch B. Cutting F. Fendrich P. Graff J. Liebetanz M. Zurini W. Jahnke (2003) J. Biomol. NMR 26 367–372 Occurrence Handle1:CAS:528:DC%2BD3sXktlagt74%3D Occurrence Handle12815263
H.A. van den Burg P.J.G.M. de Wit J. Verpoort (2001) J. Biomol. NMR 20 251–261 Occurrence Handle1:CAS:528:DC%2BD3MXmsFKnsbo%3D Occurrence Handle11519748
R.A. Venters C. Huang B.T. Farmer R. Trolard L.D. Spicer C.A. Fierke (1995) J. Biomol. NMR 5 339–344 Occurrence Handle1:CAS:528:DyaK2MXnt1ajtrw%3D Occurrence Handle7647552
S.A. Weiss G.C. Smith S.S. Kalter J.L. Vaughn (1981) In Vitro 17 495–502 Occurrence Handle1:CAS:528:DyaL3MXltFahsLY%3D
M.J. Wood E.A. Komives (1999) J. Biomol. NMR 13 149–159 Occurrence Handle1:CAS:528:DyaK1MXhvVOgsbw%3D Occurrence Handle10070756
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Strauss, A., Bitsch, F., Fendrich, G. et al. Efficient uniform isotope labeling of Abl kinase expressed in Baculovirus-infected insect cells. J Biomol NMR 31, 343–349 (2005). https://doi.org/10.1007/s10858-005-2451-3
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DOI: https://doi.org/10.1007/s10858-005-2451-3