Sequential 1H and 15N NMR assignments and secondary structure of a recombinant anti-digoxin antibody VL domain.
Related Articles Sequential 1H and 15N NMR assignments and secondary structure of a recombinant anti-digoxin antibody VL domain.
Biochemistry. 1992 Jun 2;31(21):5033-43
Authors: Constantine KL, Goldfarb V, Wittekind M, Anthony J, Ng SC, Mueller L
A uniformly 15N-labeled recombinant light-chain variable (VL) domain from the anti-digoxin antibody 26-10 has been investigated by heteronuclear two-dimensional (2D) and three-dimensional (3D) NMR spectroscopy. Complementary homonuclear 2D NMR studies of the unlabeled VL domain were also performed. Sequence-specific assignments for 97% of the main-chain and 70% of the side-chain proton resonances have been obtained. Patterns of nuclear Overhauser effects observed in 2D NOESY, 3D NOESY-HSQC, and 3D NOESY-TOCSY-HSQC spectra afford a detailed characterization of the VL domain secondary structure in solution. The observed secondary structure--a nine-stranded antiparallel beta-barrel--corresponds to that observed crystallographically for VL domains involved in quaternary associations. The locations of slowly exchanging amide protons have been discerned from a 2D TOCSY spectrum recorded after dissolving the protein in 2H2O. Strands B, C, E, and F are found to be particularly stable. The possible consequences of these results for domain-domain interactions are discussed.
PMID: 1318076 [PubMed - indexed for MEDLINE]
Source:
PubMed