Premium
Unfolding and Conformational Distributions during Protein Precipitation
Author(s) -
Chang Stephen T.,
Tobler Scott A.,
Fernandez Erik J.
Publication year - 2001
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp010025h
Subject(s) - lysozyme , chemistry , precipitation , mass spectrometry , hydrogen bond , nuclear magnetic resonance spectroscopy , protein precipitation , protein structure , biophysics , crystallography , chromatography , biochemistry , molecule , stereochemistry , biology , organic chemistry , meteorology , physics
The association of misfolded proteins, or aggregation, is a critical problem in a number of human diseases as well during the expression, refolding, formulation, and delivery of therapeutic proteins. In this study, we investigate lysozyme precipitation with hydrogen exhange using nuclear magnetic resonance (NMR) and mass spectrometry (MS). We show that MS can reveal the presence of conformational distributions, albeit without the detailed structural information afforded by NMR. Further, we find that increases in precipitant concentration alter the structure and composition of precipitates. The selective unfolding of one portion of the protein in these precipitates is correlated with hydrogen exchange patterns observed under nonprecipitating conditions and in other studies of lysozyme.