Abstract
The rugged folding landscapes of functional proteins puts them at risk of misfolding and aggregation. Serine protease inhibitors, or serpins, are paradigms for this delicate balance between function and misfolding. Serpins exist in a metastable state that undergoes a major conformational change in order to inhibit proteases. However, conformational labiality of the native serpin fold renders them susceptible to misfolding, which underlies misfolding diseases such as α1-antitrypsin deficiency. To investigate how serpins balance function and folding, we used consensus design to create conserpin, a synthetic serpin that folds reversibly, is functional, thermostable, and polymerization resistant. Characterization of its structure, folding and dynamics suggest that consensus design has remodeled the folding landscape to reconcile competing requirements for stability and function. This approach may offer general benefits for engineering functional proteins that have risky folding landscapes, including the removal of aggregation-prone intermediates, and modifying scaffolds for use as protein therapeutics.
| Original language | English |
|---|---|
| Article number | 33958 |
| Number of pages | 14 |
| Journal | Scientific Reports |
| Volume | 6 |
| DOIs | |
| Publication status | Published - 26 Sep 2016 |
Keywords
- molecular conformation
- protein folding
- x-ray crystallography
Cite this
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Smoothing a rugged protein folding landscape by sequence-based redesign. / Porebski, Benjamin T.; Keleher, Shani; Hollins, Jeffrey J.; Nickson, Adrian A.; Marijanovic, Emilia M.; Borg, Natalie A.; Costa, Mauricio G. S.; Pearce, Mary A.; Dai, Weiwen; Zhu, Liguang; Irving, James A.; Hoke, David E.; Kass, Itamar; Whisstock, James C.; Bottomley, Stephen P.; Webb, Geoffrey I.; McGowan, Sheena; Buckle, Ashley M.
In: Scientific Reports, Vol. 6, 33958, 26.09.2016.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Smoothing a rugged protein folding landscape by sequence-based redesign
AU - Porebski, Benjamin T.
AU - Keleher, Shani
AU - Hollins, Jeffrey J.
AU - Nickson, Adrian A.
AU - Marijanovic, Emilia M.
AU - Borg, Natalie A.
AU - Costa, Mauricio G. S.
AU - Pearce, Mary A.
AU - Dai, Weiwen
AU - Zhu, Liguang
AU - Irving, James A.
AU - Hoke, David E.
AU - Kass, Itamar
AU - Whisstock, James C.
AU - Bottomley, Stephen P.
AU - Webb, Geoffrey I.
AU - McGowan, Sheena
AU - Buckle, Ashley M.
PY - 2016/9/26
Y1 - 2016/9/26
N2 - The rugged folding landscapes of functional proteins puts them at risk of misfolding and aggregation. Serine protease inhibitors, or serpins, are paradigms for this delicate balance between function and misfolding. Serpins exist in a metastable state that undergoes a major conformational change in order to inhibit proteases. However, conformational labiality of the native serpin fold renders them susceptible to misfolding, which underlies misfolding diseases such as α1-antitrypsin deficiency. To investigate how serpins balance function and folding, we used consensus design to create conserpin, a synthetic serpin that folds reversibly, is functional, thermostable, and polymerization resistant. Characterization of its structure, folding and dynamics suggest that consensus design has remodeled the folding landscape to reconcile competing requirements for stability and function. This approach may offer general benefits for engineering functional proteins that have risky folding landscapes, including the removal of aggregation-prone intermediates, and modifying scaffolds for use as protein therapeutics.
AB - The rugged folding landscapes of functional proteins puts them at risk of misfolding and aggregation. Serine protease inhibitors, or serpins, are paradigms for this delicate balance between function and misfolding. Serpins exist in a metastable state that undergoes a major conformational change in order to inhibit proteases. However, conformational labiality of the native serpin fold renders them susceptible to misfolding, which underlies misfolding diseases such as α1-antitrypsin deficiency. To investigate how serpins balance function and folding, we used consensus design to create conserpin, a synthetic serpin that folds reversibly, is functional, thermostable, and polymerization resistant. Characterization of its structure, folding and dynamics suggest that consensus design has remodeled the folding landscape to reconcile competing requirements for stability and function. This approach may offer general benefits for engineering functional proteins that have risky folding landscapes, including the removal of aggregation-prone intermediates, and modifying scaffolds for use as protein therapeutics.
KW - molecular conformation
KW - protein folding
KW - x-ray crystallography
UR - http://www.scopus.com/inward/record.url?scp=84988927096&partnerID=8YFLogxK
U2 - 10.1038/srep33958
DO - 10.1038/srep33958
M3 - Article
AN - SCOPUS:84988927096
VL - 6
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
M1 - 33958
ER -