Smoothing a rugged protein folding landscape by sequence-based redesign

Benjamin T. Porebski, Shani Keleher, Jeffrey J. Hollins, Adrian A. Nickson, Emilia M. Marijanovic, Natalie A. Borg, Mauricio G. S. Costa, Mary A. Pearce, Weiwen Dai, Liguang Zhu, James A. Irving, David E. Hoke, Itamar Kass, James C. Whisstock, Stephen P. Bottomley, Geoffrey I. Webb, Sheena McGowan, Ashley M. Buckle

Research output: Contribution to journalArticleResearchpeer-review

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 languageEnglish
Article number33958
Number of pages14
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 26 Sep 2016

Keywords

  • molecular conformation
  • protein folding
  • x-ray crystallography

Cite this

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. / Smoothing a rugged protein folding landscape by sequence-based redesign. In: Scientific Reports. 2016 ; Vol. 6.
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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.",
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author = "Porebski, {Benjamin T.} and Shani Keleher and Hollins, {Jeffrey J.} and Nickson, {Adrian A.} and Marijanovic, {Emilia M.} and Borg, {Natalie A.} and Costa, {Mauricio G. S.} and Pearce, {Mary A.} and Weiwen Dai and Liguang Zhu and Irving, {James A.} and Hoke, {David E.} and Itamar Kass and Whisstock, {James C.} and Bottomley, {Stephen P.} and Webb, {Geoffrey I.} and Sheena McGowan and Buckle, {Ashley M.}",
year = "2016",
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Porebski, BT, Keleher, S, Hollins, JJ, Nickson, AA, Marijanovic, EM, Borg, NA, Costa, MGS, Pearce, MA, Dai, W, Zhu, L, Irving, JA, Hoke, DE, Kass, I, Whisstock, JC, Bottomley, SP, Webb, GI, McGowan, S & Buckle, AM 2016, 'Smoothing a rugged protein folding landscape by sequence-based redesign' Scientific Reports, vol. 6, 33958. https://doi.org/10.1038/srep33958

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 journalArticleResearchpeer-review

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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.

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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.

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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.

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