ISSN:
1573-5001
Keywords:
Antibody domain
;
Chemical shift
;
Side-chain assignments
;
Triple resonance
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Chemistry and Pharmacology
Notes:
Summary 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, Hα and Cα resonances. These data confirm and extend HN, N and Hα 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 Hα and Cα 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 Cα and Cβ 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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00242474
