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Comparison of the amide proton exchange behavior of the rapidly formed folding intermediate and the native state of an antibody scFv fragment
Author(s) -
Freund Cristian,
Gehrig Peter,
Holak Tad A,
Plückthun Andreas
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00306-2
Subject(s) - chemistry , amide , folding (dsp implementation) , native state , intermediate state , crystallography , protein folding , stereochemistry , proton , biochemistry , physics , atomic physics , quantum mechanics , electrical engineering , engineering
We have investigated the stability of backbone amide protons of the intermediate and the native state of the scFv fragment of an antibody. Stopped flow experiments analyzed by MS and NMR detected the formation of an exchange protected intermediate within the deadtime of the stopped flow apparatus (17 ms). H/D exchange rates of the native protein identified a number of very stable backbone amide protons in the V L and the V H domains. In the V L domain, this slowly exchanging core of the scFv fragment is similar to the folding core of the intermediate, while the V H domain possesses a great number of very stable amide protons which are not stabilized to a significant degree in the folding intermediate of the scFv fragment.