Small molecule diselenide additives for in vitro oxidative protein folding

2016 ◽  
Vol 52 (16) ◽  
pp. 3336-3339 ◽  
Author(s):  
Post Sai Reddy ◽  
Norman Metanis
Keyword(s):  
Protein Folding ◽  
Small Molecule ◽  
Oxidative Folding ◽  
Oxidative Protein Folding

Small molecule diselenides were prepared and found to enhance thein vitrooxidative folding of disulfide-rich protein.

scholarly journals Diselenide Crosslinks for Enhanced and Simplified Oxidative Protein Folding

2020 ◽  
Author(s):  
Reem Mousa ◽  
Taghreed Hidmi ◽  
Sergei Pomyalov ◽  
Shifra Lansky ◽  
Lareen Khouri ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Protein Folding ◽  
Disulfide Bonds ◽  
Crystal Structure Analysis ◽  
Folding Rate ◽  
Oxidative Folding ◽  
Functional Studies ◽  
Oxidative Protein Folding ◽  
And Function

<p>The oxidative folding of proteins has been studied for over sixty years, providing critical insight into protein folding mechanisms. A well-known folding model for many disulfide-rich proteins is that of hirudin. Hirudin, the most potent natural inhibitor of thrombin, is a 65-residue protein with three disulfide bonds, and folds through plagued pathway that involve highly heterogeneous intermediates and scrambled isomers. The formation of scrambled species is known to limit the rate and efficiency of <i>in vitro</i> oxidative folding of many proteins.</p><p>In the current manuscript we describe our recent work, intended to overcome the limitations of scrambled isomers formation during oxidative protein folding. In this research we deeply investigate the utility of introducing diselenide bridges at the three native disulfide crosslinks as well as at a non-native position on hirudin’s folding, structure and function. Our studies demonstrated that, regardless of the specific positions of these substitutions, the diselenide crosslinks enhanced the folding rate and yield of the hirudin analogs, while reducing the complexity and heterogeneity of the process, and reducing the formation of scrambled isomers.</p><p>A parallel, equally important, objective of our study was to test if diselenide substitutions have structural and functional effects. Crystal structure analysis as well as functional studies indicated that diselenide crosslinks maintained the overall structure of the protein without causing major changes in function and structure. To substantiate these conclusions, we provide inhibition studies and high-resolution crystal structure of the wild-type hirudin and its seleno-analogs. </p>Taken together, we believe that the choice of hirudin as the model in this study has implications beyond its specific folding mechanism, and will serve as a useful methodology for the <i>in vitro</i> oxidative folding of many complex disulfide-rich proteins.

scholarly journals Diselenide Crosslinks for Enhanced and Simplified Oxidative Protein Folding

2020 ◽  
Author(s):  
Reem Mousa ◽  
Taghreed Hidmi ◽  
Sergei Pomyalov ◽  
Shifra Lansky ◽  
Lareen Khouri ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Protein Folding ◽  
Disulfide Bonds ◽  
Crystal Structure Analysis ◽  
Folding Rate ◽  
Oxidative Folding ◽  
Functional Studies ◽  
Oxidative Protein Folding ◽  
And Function

<p>The oxidative folding of proteins has been studied for over sixty years, providing critical insight into protein folding mechanisms. A well-known folding model for many disulfide-rich proteins is that of hirudin. Hirudin, the most potent natural inhibitor of thrombin, is a 65-residue protein with three disulfide bonds, and folds through plagued pathway that involve highly heterogeneous intermediates and scrambled isomers. The formation of scrambled species is known to limit the rate and efficiency of <i>in vitro</i> oxidative folding of many proteins.</p><p>In the current manuscript we describe our recent work, intended to overcome the limitations of scrambled isomers formation during oxidative protein folding. In this research we deeply investigate the utility of introducing diselenide bridges at the three native disulfide crosslinks as well as at a non-native position on hirudin’s folding, structure and function. Our studies demonstrated that, regardless of the specific positions of these substitutions, the diselenide crosslinks enhanced the folding rate and yield of the hirudin analogs, while reducing the complexity and heterogeneity of the process, and reducing the formation of scrambled isomers.</p><p>A parallel, equally important, objective of our study was to test if diselenide substitutions have structural and functional effects. Crystal structure analysis as well as functional studies indicated that diselenide crosslinks maintained the overall structure of the protein without causing major changes in function and structure. To substantiate these conclusions, we provide inhibition studies and high-resolution crystal structure of the wild-type hirudin and its seleno-analogs. </p>Taken together, we believe that the choice of hirudin as the model in this study has implications beyond its specific folding mechanism, and will serve as a useful methodology for the <i>in vitro</i> oxidative folding of many complex disulfide-rich proteins.

Catalysis of Oxidative Protein Folding by Small-Molecule Diselenides†

Biochemistry ◽  
2008 ◽  
Vol 47 (27) ◽  
pp. 6985-6987 ◽  
Author(s):  
Joris Beld ◽  
Kenneth J. Woycechowsky ◽  
Donald Hilvert
Keyword(s):  
Protein Folding ◽  
Small Molecule ◽  
Oxidative Protein Folding

scholarly journals Redox-Active Cyclic Bis(cysteinyl)peptides as Catalysts for In Vitro Oxidative Protein Folding

2002 ◽  
Vol 9 (6) ◽  
pp. 731-740 ◽  
Author(s):  
Chiara Cabrele ◽  
Stella Fiori ◽  
Stefano Pegoraro ◽  
Luis Moroder
Keyword(s):  
Protein Folding ◽  
Oxidative Protein Folding ◽  
Redox Active

scholarly journals Quantitative Analyses of the Yeast Oxidative Protein Folding Pathway In Vitro and In Vivo

2019 ◽  
Vol 31 (4) ◽  
pp. 261-274 ◽  
Author(s):  
Dave M. Beal ◽  
Emma L. Bastow ◽  
Gemma L. Staniforth ◽  
Tobias von der Haar ◽  
Robert B. Freedman ◽  
...  
Keyword(s):  
Protein Folding ◽  
Folding Pathway ◽  
Oxidative Protein Folding ◽  
Quantitative Analyses

scholarly journals Oxidative Folding of Proteins: The “Smoking Gun” of Glutathione

2021 ◽  
Vol 22 (18) ◽  
pp. 10148
Author(s):  
Alessio Bocedi ◽  
Giada Cattani ◽  
Giorgia Gambardella ◽  
Linda Schulte ◽  
Harald Schwalbe ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Protein Folding ◽  
Criminal Trial ◽  
Low Molecular Weight ◽  
Oxidative Folding ◽  
Oxidative Protein Folding

Glutathione has long been suspected to be the primary low molecular weight compound present in all cells promoting the oxidative protein folding, but twenty years ago it was found “not guilty”. Now, new surprising evidence repeats its request to be the “smoking gun” which reopens the criminal trial revealing the crucial involvement of this tripeptide.

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