Oxidative Protein Folding: From the Test Tube to In Vivo Insights

2008 ◽  
Vol 10 (1) ◽  
pp. 51-54 ◽  
Author(s):  
Salvador Ventura
Keyword(s):  
Protein Folding ◽  
Test Tube ◽  
Oxidative Protein Folding

scholarly journals Endoplasmic reticulum oxidoreductin (ERO) provides resilience against reductive stress and hypoxic conditions by mediating luminal redox dynamics

2021 ◽  
Author(s):  
Jose Manuel Ugalde ◽  
Isabel Aller ◽  
Lika Kudrjasova ◽  
Romy Schmidt ◽  
Michelle Schloesser ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Folding ◽  
Redox Homeostasis ◽  
Midpoint Potential ◽  
Oxidative Protein Folding ◽  
Reductive Stress ◽  
Hypoxic Conditions ◽  
Glutathione Redox ◽  
Redox Dynamics

Oxidative protein folding in the endoplasmic reticulum (ER) depends on the coordinated action of protein disulfide isomerases and ER oxidoreductins (EROs). Strict dependence of ERO activity on molecular oxygen as the final electron acceptor implies that oxidative protein folding and other ER processes are severely compromised under hypoxia. While many key players involved in oxidative protein folding are known, our understanding of how redox homeostasis in the ER is maintained and how EROs, the Cys residues of nascent proteins, and the luminal glutathione redox buffer interact is limited. Here, we isolated viable ero1 ero2 double mutants largely deficient in ERO activity, which rendered the mutants highly sensitive to reductive stress and hypoxia. To elucidate the specific redox dynamics in the ER lumen in vivo, we expressed the glutathione redox potential (EGSH) sensor Grx1-roGFP2iL-HDEL with a midpoint potential of -240 mV in the ER of Arabidopsis plants. We found EGSH values of -241 mV in wild-type plants, which is less oxidizing than previously estimated. In the ero1 ero2 mutants, luminal EGSH was reduced further to -253 mV. Recovery to reductive ER stress, as induced by acute exposure to dithiothreitol, was delayed in ero1 ero2 mutants. The characteristic signature of EGSH dynamics in the ER lumen triggered by hypoxia was affected in the ero1 ero2 mutant reflecting a disrupted balance of reductive and oxidizing inputs, including nascent polypeptides and glutathione entry. The ER redox dynamics can now be dissected in vivo, revealing a central role of EROs as major redox integrators to promote luminal redox homeostasis.

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

Small-Molecule Diselenides Catalyze Oxidative Protein Folding in Vivo

2010 ◽  
Vol 5 (2) ◽  
pp. 177-182 ◽  
Author(s):  
Joris Beld ◽  
Kenneth J. Woycechowsky ◽  
Donald Hilvert
Keyword(s):  
Protein Folding ◽  
Small Molecule ◽  
Oxidative Protein Folding

Silent mutations affect in vivo protein folding in Escherichia coli

2002 ◽  
Vol 293 (1) ◽  
pp. 537-541 ◽  
Author(s):  
Patricia Cortazzo ◽  
Carlos Cerveñansky ◽  
Mónica Marı́n ◽  
Claude Reiss ◽  
Ricardo Ehrlich ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Folding

scholarly journals Conjugate of Thiol and Guanidyl Units with Oligoethylene Glycol Linkage for Manipulation of Oxidative Protein Folding

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 879
Author(s):  
Shunsuke Okada ◽  
Motonori Matsusaki ◽  
Masaki Okumura ◽  
Takahiro Muraoka
Keyword(s):  
Protein Folding ◽  
Active Sites ◽  
Biological Process ◽  
Thiol Group ◽  
Native Conformation ◽  
Thiol Compounds ◽  
Oxidative Protein Folding ◽  
Redox Active ◽  
A Cell ◽  
Protein Disulfide

Oxidative protein folding is a biological process to obtain a native conformation of a protein through disulfide-bond formation between cysteine residues. In a cell, disulfide-catalysts such as protein disulfide isomerase promote the oxidative protein folding. Inspired by the active sites of the disulfide-catalysts, synthetic redox-active thiol compounds have been developed, which have shown significant promotion of the folding processes. In our previous study, coupling effects of a thiol group and guanidyl unit on the folding promotion were reported. Herein, we investigated the influences of a spacer between the thiol group and guanidyl unit. A conjugate between thiol and guanidyl units with a diethylene glycol spacer (GdnDEG-SH) showed lower folding promotion effect compared to the thiol–guanidyl conjugate without the spacer (GdnSH). Lower acidity and a more reductive property of the thiol group of GdnDEG-SH compared to those of GdnSH likely resulted in the reduced efficiency of the folding promotion. Thus, the spacer between the thiol and guanidyl groups is critical for the promotion of oxidative protein folding.

scholarly journals Regulation of plant ER oxidoreductin 1 (ERO1) activity for efficient oxidative protein folding

2019 ◽  
Vol 294 (49) ◽  
pp. 18820-18835 ◽  
Author(s):  
Motonori Matsusaki ◽  
Aya Okuda ◽  
Koichi Matsuo ◽  
Kunihiko Gekko ◽  
Taro Masuda ◽  
...  
Keyword(s):  
Protein Folding ◽  
Oxidative Protein Folding

Principles of chaperone-mediated protein folding

1995 ◽  
Vol 348 (1323) ◽  
pp. 107-112 ◽  
Keyword(s):  
Protein Folding ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Molecular Chaperones ◽  
Cellular Protein ◽  
Chaperone Proteins ◽  
Shock Proteins ◽  
Polypeptide Chains

The recent discovery of molecular chaperones and their functions has changed dramatically our view of the processes underlying the folding of proteins in vivo . Rather than folding spontaneously, most newly synthesized polypeptide chains seem to acquire their native conformations in a reaction mediated by chaperone proteins. Different classes of molecular chaperones, such as the members of the Hsp70 and Hsp60 families of heat-shock proteins, cooperate in a coordinated pathway of cellular protein folding.

scholarly journals Protein Folding: Search for Basic Physical Models

2003 ◽  
Vol 3 ◽  
pp. 623-635 ◽  
Author(s):  
Ivan Y. Torshin ◽  
Robert W. Harrison
Keyword(s):  
Protein Folding ◽  
Tertiary Structure ◽  
Three Dimensional ◽  
Physical Models ◽  
Dimensional Structure ◽  
Computational Techniques ◽  
Linear Sequence ◽  
Physical Forces

How a unique three-dimensional structure is rapidly formed from the linear sequence of a polypeptide is one of the important questions in contemporary science. Apart from biological context ofin vivoprotein folding (which has been studied only for a few proteins), the roles of the fundamental physical forces in thein vitrofolding remain largely unstudied. Despite a degree of success in using descriptions based on statistical and/or thermodynamic approaches, few of the current models explicitly include more basic physical forces (such as electrostatics and Van Der Waals forces). Moreover, the present-day models rarely take into account that the protein folding is, essentially, a rapid process that produces a highly specific architecture. This review considers several physical models that may provide more direct links between sequence and tertiary structure in terms of the physical forces. In particular, elaboration of such simple models is likely to produce extremely effective computational techniques with value for modern genomics.

scholarly journals Computational Protein Stabilization Can Affect Folding Energy Landscapes and Lead to Domain-Swapped Dimers

2021 ◽  
Author(s):  
Klara Markova ◽  
Antonin Kunka ◽  
Klaudia Chmelova ◽  
Martin Havlasek ◽  
Petra Babkova ◽  
...  
Keyword(s):  
Protein Folding ◽  
Protein Design ◽  
Energy Landscape ◽  
Single Point ◽  
Three Dimensional ◽  
Protein Stabilization ◽  
Dimensional Structure ◽  
Evolutionary Analysis ◽  
Folding Energy

<p>The functionality of a protein depends on its unique three-dimensional structure, which is a result of the folding process when the nascent polypeptide follows a funnel-like energy landscape to reach a global energy minimum. Computer-encoded algorithms are increasingly employed to stabilize native proteins for use in research and biotechnology applications. Here, we reveal a unique example where the computational stabilization of a monomeric α/β-hydrolase enzyme (<i>T</i><sub>m</sub> = 73.5°C; Δ<i>T</i><sub>m</sub> > 23°C) affected the protein folding energy landscape. Introduction of eleven single-point stabilizing mutations based on force field calculations and evolutionary analysis yielded catalytically active domain-swapped intermediates trapped in local energy minima. Crystallographic structures revealed that these stabilizing mutations target cryptic hinge regions and newly introduced secondary interfaces, where they make extensive non-covalent interactions between the intertwined misfolded protomers. The existence of domain-swapped dimers in a solution is further confirmed experimentally by data obtained from SAXS and crosslinking mass spectrometry. Unfolding experiments showed that the domain-swapped dimers can be irreversibly converted into native-like monomers, suggesting that the domain-swapping occurs exclusively <i>in vivo</i>. Our findings uncovered hidden protein-folding consequences of computational protein design, which need to be taken into account when applying a rational stabilization to proteins of biological and pharmaceutical interest.</p>

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