TY - JOUR
T1 - Combining Structural Aggregation Propensity and Stability Predictions to Redesign Protein Solubility
AU - Gil-Garcia, Marcos
AU - Banó-Polo, Manuel
AU - Varejao, Nathalia
AU - Jamroz, Michal
AU - Kuriata, Aleksander
AU - Díaz-Caballero, Marta
AU - Lascorz, Jara
AU - Morel, Bertrand
AU - Navarro, Susanna
AU - Reverter, David
AU - Kmiecik, Sebastian
AU - Ventura, Salvador
PY - 2018/9/4
Y1 - 2018/9/4
N2 - Copyright © 2018 American Chemical Society. The aggregation propensity of each particular protein seems to be shaped by evolution according to its natural abundance in the cell. The production and downstream processing of recombinant polypeptides implies attaining concentrations that are orders of magnitude above their natural levels, often resulting in their aggregation; a phenomenon that precludes the marketing of many globular proteins for biomedical or biotechnological applications. Therefore, there is a huge interest in methods aimed to increase the proteins solubility above their natural limits. Here, we demonstrate that an updated version of our AGGRESCAN 3D structural aggregation predictor, that now takes into account protein stability, allows for designing mutations at specific positions in the structure that improve the solubility of proteins without compromising their conformation. Using this approach, we have designed a highly soluble variant of the green fluorescent protein and a human single-domain VH antibody displaying significantly reduced aggregation propensity. Overall, our data indicate that the solubility of unrelated proteins can be easily tuned by in silico-designed nondestabilizing amino acid changes at their surfaces.
AB - Copyright © 2018 American Chemical Society. The aggregation propensity of each particular protein seems to be shaped by evolution according to its natural abundance in the cell. The production and downstream processing of recombinant polypeptides implies attaining concentrations that are orders of magnitude above their natural levels, often resulting in their aggregation; a phenomenon that precludes the marketing of many globular proteins for biomedical or biotechnological applications. Therefore, there is a huge interest in methods aimed to increase the proteins solubility above their natural limits. Here, we demonstrate that an updated version of our AGGRESCAN 3D structural aggregation predictor, that now takes into account protein stability, allows for designing mutations at specific positions in the structure that improve the solubility of proteins without compromising their conformation. Using this approach, we have designed a highly soluble variant of the green fluorescent protein and a human single-domain VH antibody displaying significantly reduced aggregation propensity. Overall, our data indicate that the solubility of unrelated proteins can be easily tuned by in silico-designed nondestabilizing amino acid changes at their surfaces.
KW - Aβ peptide
KW - green fluorescent protein
KW - protein aggregation
KW - protein stability
KW - protein structure
KW - single-domain antibodies
U2 - 10.1021/acs.molpharmaceut.8b00341
DO - 10.1021/acs.molpharmaceut.8b00341
M3 - Article
C2 - 30036481
VL - 15
SP - 3846
EP - 3859
JO - Molecular Pharmaceutics
JF - Molecular Pharmaceutics
SN - 1543-8384
ER -