Constantine K L, Goldfarb V, Wittekind M, Friedrichs M S, Anthony J, Ng S C, Mueller L
Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ 08543.
J Biomol NMR. 1993 Jan;3(1):41-54. doi: 10.1007/BF00242474.
Extensive 1H and 13C assignments have been obtained for the aliphatic resonances of a uniformly 13C- and 15N-labeled recombinant VL domain from the anti-digoxin antibody 26-10. Four-dimensional triple resonance NMR data acquired with the HNCAHA and HN(CO)CAHA pulse sequences [Kay et al. (1992) J. Magn. Reson., 98, 443-450] afforded assignments for the backbone HN, N, H alpha and C alpha resonances. These data confirm and extend HN, N and H alpha assignments derived previously from three-dimensional 1H-15N NMR studies of uniformly 15N-labeled VL domain [Constantine et al. (1992), Biochemistry, 31, 5033-5043]. The identified H alpha and C alpha resonances provided a starting point for assigning the side-chain aliphatic 1H and 13C resonances using three-dimensional HCCH-COSY and HCCH-TOCSY experiments [Clore et al. (1990), Biochemistry, 29, 8172-8184]. The C alpha and C beta chemical shifts are correlated with the VL domain secondary structure. The extensive set of side-chain assignments obtained will allow a detailed comparison to be made between the solution structure of the isolated VL domain and the X-ray structure of the VL domain within the 26-10 Fab.
已获得来自抗地高辛抗体26 - 10的均匀13C和15N标记的重组VL结构域脂肪族共振的广泛1H和13C归属。用HNCAHA和HN(CO)CAHA脉冲序列[Kay等人，(1992)《磁共振杂志》，98, 443 - 450]采集的四维三重共振NMR数据给出了主链HN、N、Hα和Cα共振的归属。这些数据证实并扩展了先前从均匀15N标记的VL结构域的三维1H - 15N NMR研究中得出的HN、N和Hα归属[Constantine等人，(1992)，《生物化学》，31, 5033 - 5043]。鉴定出的Hα和Cα共振为使用三维HCCH - COSY和HCCH - TOCSY实验[Clore等人，(1990)，《生物化学》，29, 8172 - 8184]归属侧链脂肪族1H和13C共振提供了起点。Cα和Cβ化学位移与VL结构域二级结构相关。所获得的广泛侧链归属将使得能够对分离的VL结构域的溶液结构与26 - 10 Fab内VL结构域的X射线结构进行详细比较。
