scholarly journals Ero1–PDI interactions, the response to redox flux and the implications for disulfide bond formation in the mammalian endoplasmic reticulum

2013 ◽  
Vol 368 (1617) ◽  
pp. 20110403 ◽  
Author(s):  
Adam M. Benham ◽  
Marcel van Lith ◽  
Roberto Sitia ◽  
Ineke Braakman
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Folding ◽  
Disulfide Bond ◽  
Disulfide Bonds ◽  
Secretory Pathway ◽  
Protein Complexes ◽  
Bond Formation ◽  
Disulfide Bond Formation ◽  
Terminal Electron Acceptor ◽  
Post Translational Modifications

The protein folding machinery of the endoplasmic reticulum (ER) ensures that proteins entering the eukaryotic secretory pathway acquire appropriate post-translational modifications and reach a stably folded state. An important component of this protein folding process is the supply of disulfide bonds. These are introduced into client proteins by ER resident oxidoreductases, including ER oxidoreductin 1 (Ero1). Ero1 is usually considered to function in a linear pathway, by ‘donating’ a disulfide bond to protein disulfide isomerase (PDI) and receiving electrons that are passed on to the terminal electron acceptor molecular oxygen. PDI engages with a range of clients as the direct catalyst of disulfide bond formation, isomerization or reduction. In this paper, we will consider the interactions of Ero1 with PDI family proteins and chaperones, highlighting the effect that redox flux has on Ero1 partnerships. In addition, we will discuss whether higher order protein complexes play a role in Ero1 function.

scholarly journals Two phases of disulfide bond formation have differing requirements for oxygen

2013 ◽  
Vol 203 (4) ◽  
pp. 615-627 ◽  
Author(s):  
Marianne Koritzinsky ◽  
Fiana Levitin ◽  
Twan van den Beucken ◽  
Ryan A. Rumantir ◽  
Nicholas J. Harding ◽  
...  
Keyword(s):  
Er Stress ◽  
Disulfide Bond ◽  
Electron Acceptor ◽  
Disulfide Bonds ◽  
Mammalian Cells ◽  
Secretory Pathway ◽  
Bond Formation ◽  
Disulfide Bond Formation ◽  
Terminal Electron Acceptor ◽  
Two Phases

Most proteins destined for the extracellular space require disulfide bonds for folding and stability. Disulfide bonds are introduced co- and post-translationally in endoplasmic reticulum (ER) cargo in a redox relay that requires a terminal electron acceptor. Oxygen can serve as the electron acceptor in vitro, but its role in vivo remains unknown. Hypoxia causes ER stress, suggesting a role for oxygen in protein folding. Here we demonstrate the existence of two phases of disulfide bond formation in living mammalian cells, with differential requirements for oxygen. Disulfide bonds introduced rapidly during protein synthesis can occur without oxygen, whereas those introduced during post-translational folding or isomerization are oxygen dependent. Other protein maturation processes in the secretory pathway, including ER-localized N-linked glycosylation, glycan trimming, Golgi-localized complex glycosylation, and protein transport, occur independently of oxygen availability. These results suggest that an alternative electron acceptor is available transiently during an initial phase of disulfide bond formation and that post-translational oxygen-dependent disulfide bond formation causes hypoxia-induced ER stress.

355 HYPOXIA INHIBITS DISULFIDE BOND FORMATION AND PROTEIN FOLDING IN THE ENDOPLASMIC RETICULUM

2012 ◽  
Vol 102 ◽  
pp. S185-S186
Author(s):  
M. Koritzinsky ◽  
T. Van den Beucken ◽  
K. Chu ◽  
P.C. Boutros ◽  
I. Braakman ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Folding ◽  
Disulfide Bond ◽  
Bond Formation ◽  
Disulfide Bond Formation

scholarly journals From recordings of disulfide isomerases in action to reversal of maladaptive endoplasmic reticulum stress responses: proceedings on the ER & Redox Club Meeting held in Venice, April 2015

2015 ◽  
Vol 2 (1) ◽  
Author(s):  
Eelco van Anken
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Folding ◽  
Disulfide Bond ◽  
Stress Responses ◽  
Bond Formation ◽  
Genetic Defect ◽  
Cell Protein ◽  
Disulfide Bond Formation ◽  
Club Meeting ◽  
Client Proteins

AbstractThe endoplasmic reticulum (ER) interacts and cooperates with other organelles as a central hub in cellular homeostasis. In particular, the ER is the first station along the secretory pathway, where client proteins fold and assemble before they travel to their final destination elsewhere in the endomembrane system or outside the cell. Protein folding and disulfide bond formation go hand in hand in the ER, a task that is achieved with the help of ER-resident chaperones and other folding factors, including oxidoreductases that catalyze disulfide bond formation. Yet, when their combined effort is in vain, client proteins that fail to fold are disposed of through ER-associated degradation (ERAD). The ER folding and ERAD machineries can be boosted through the unfolded protein response (UPR) if required. Still, protein folding in the ER may consistently fail when proteins are mutated due to a genetic defect, which, ultimately, can lead to disease. Novel developments in all these fields of study and how new insights ultimately can be exploited for clinical or biotechnological purposes were highlighted in a rich variety of presentations at the ER & Redox Club Meeting that was held in Venice from 15 to 17 April 2015. As such, the meeting provided the participants an excellent opportunity to mingle and discuss key advancements and outstanding questions on ER function in health and disease.

scholarly journals Mechanisms of Disulfide Bond Formation in Nascent Polypeptides Entering the Secretory Pathway

Cells ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1994 ◽  
Author(s):  
Philip J. Robinson ◽  
Neil J. Bulleid
Keyword(s):  
Disulfide Bond ◽  
Disulfide Bonds ◽  
Secretory Pathway ◽  
Bond Formation ◽  
Complex Problem ◽  
Structure Stability ◽  
Disulfide Bond Formation ◽  
Polypeptide Folding ◽  
Domains Of Life ◽  
And Function

Disulfide bonds are an abundant feature of proteins across all domains of life that are important for structure, stability, and function. In eukaryotic cells, a major site of disulfide bond formation is the endoplasmic reticulum (ER). How cysteines correctly pair during polypeptide folding to form the native disulfide bond pattern is a complex problem that is not fully understood. In this paper, the evidence for different folding mechanisms involved in ER-localised disulfide bond formation is reviewed with emphasis on events that occur during ER entry. Disulfide formation in nascent polypeptides is discussed with focus on (i) its mechanistic relationship with conformational folding, (ii) evidence for its occurrence at the co-translational stage during ER entry, and (iii) the role of protein disulfide isomerase (PDI) family members. This review highlights the complex array of cellular processes that influence disulfide bond formation and identifies key questions that need to be addressed to further understand this fundamental process.

scholarly journals PDI-Regulated Disulfide Bond Formation in Protein Folding and Biomolecular Assembly

Molecules ◽  
2020 ◽  
Vol 26 (1) ◽  
pp. 171
Author(s):  
Jiahui Fu ◽  
Jihui Gao ◽  
Zhongxin Liang ◽  
Dong Yang
Keyword(s):  
Protein Folding ◽  
Disulfide Bond ◽  
Disulfide Bonds ◽  
Protein Disulfide Isomerase ◽  
Bond Formation ◽  
Molecular Assembly ◽  
Disulfide Bond Formation ◽  
Disulfide Isomerase ◽  
Biomolecular Assembly ◽  
Protein Disulfide

Disulfide bonds play a pivotal role in maintaining the natural structures of proteins to ensure their performance of normal biological functions. Moreover, biological molecular assembly, such as the gluten network, is also largely dependent on the intermolecular crosslinking via disulfide bonds. In eukaryotes, the formation and rearrangement of most intra- and intermolecular disulfide bonds in the endoplasmic reticulum (ER) are mediated by protein disulfide isomerases (PDIs), which consist of multiple thioredoxin-like domains. These domains assist correct folding of proteins, as well as effectively prevent the aggregation of misfolded ones. Protein misfolding often leads to the formation of pathological protein aggregations that cause many diseases. On the other hand, glutenin aggregation and subsequent crosslinking are required for the formation of a rheologically dominating gluten network. Herein, the mechanism of PDI-regulated disulfide bond formation is important for understanding not only protein folding and associated diseases, but also the formation of functional biomolecular assembly. This review systematically illustrated the process of human protein disulfide isomerase (hPDI) mediated disulfide bond formation and complemented this with the current mechanism of wheat protein disulfide isomerase (wPDI) catalyzed formation of gluten networks.

scholarly journals In situ detection of protein interactions for recombinant therapeutic enzymes

2020 ◽  
Author(s):  
Mojtaba Samoudi ◽  
Chih-Chung Kuo ◽  
Caressa M. Robinson ◽  
Km Shams-Ud-Doha ◽  
Song-Min Schinn ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Recombinant Proteins ◽  
Disulfide Bond ◽  
Disulfide Bonds ◽  
Secretory Pathway ◽  
Molecular Mechanisms ◽  
Therapeutic Potential ◽  
Bond Formation ◽  
Disulfide Bond Formation ◽  
Ppi Networks

AbstractDespite their therapeutic potential, many protein drugs remain inaccessible to patients since they are difficult to secrete. Each recombinant protein has unique physicochemical properties and requires different machinery for proper folding, assembly, and post-translational modifications (PTMs). Here we aimed to identify the machinery supporting recombinant protein secretion by measuring the protein-protein interaction (PPI) networks of four different recombinant proteins (SERPINA1, SERPINC1, SERPING1 and SeAP) with various PTMs and structural motifs using the proximity-dependent biotin identification (BioID) method. We identified PPIs associated with specific features of the secreted proteins using a Bayesian statistical model, and found proteins involved in protein folding, disulfide bond formation and N-glycosylation were positively correlated with the corresponding features of the four model proteins. Among others, oxidative folding enzymes showed the strongest association with disulfide bond formation, supporting their critical roles in proper folding and maintaining the ER stability. Knockdown of disulfide-isomerase PDIA4, a measured interactor with significance for SERPINC1 but not SERPINA1, led to the decreased secretion of SERPINC1, which relies on its extensive disulfide bonds, compared to SERPINA1, which has no disulfide bonds. Proximity-dependent labeling successfully identified the transient interactions supporting synthesis of secreted recombinant proteins and refined our understanding of key molecular mechanisms of the secretory pathway during recombinant protein production.

scholarly journals Protein folding guides disulfide bond formation

2015 ◽  
Vol 112 (36) ◽  
pp. 11241-11246 ◽  
Author(s):  
Meng Qin ◽  
Wei Wang ◽  
D. Thirumalai
Keyword(s):  
Protein Folding ◽  
Disulfide Bond ◽  
Disulfide Bonds ◽  
Broad Class ◽  
Bond Formation ◽  
Coarse Grained ◽  
Disulfide Bond Formation ◽  
Native State ◽  
Experimental Findings ◽  
Β Sheet

The Anfinsen principle that the protein sequence uniquely determines its structure is based on experiments on oxidative refolding of a protein with disulfide bonds. The problem of how protein folding drives disulfide bond formation is poorly understood. Here, we have solved this long-standing problem by creating a general method for implementing the chemistry of disulfide bond formation and rupture in coarse-grained molecular simulations. As a case study, we investigate the oxidative folding of bovine pancreatic trypsin inhibitor (BPTI). After confirming the experimental findings that the multiple routes to the folded state contain a network of states dominated by native disulfides, we show that the entropically unfavorable native single disulfide [14–38] between Cys14 and Cys38 forms only after polypeptide chain collapse and complete structuring of the central core of the protein containing an antiparallel β-sheet. Subsequent assembly, resulting in native two-disulfide bonds and the folded state, involves substantial unfolding of the protein and transient population of nonnative structures. The rate of [14–38] formation increases as the β-sheet stability increases. The flux to the native state, through a network of kinetically connected native-like intermediates, changes dramatically by altering the redox conditions. Disulfide bond formation between Cys residues not present in the native state are relevant only on the time scale of collapse of BPTI. The finding that formation of specific collapsed native-like structures guides efficient folding is applicable to a broad class of single-domain proteins, including enzyme-catalyzed disulfide proteins.

scholarly journals A Relaxed Specificity in Interchain Disulfide Bond Formation Characterizes the Assembly of a Low-Molecular-Weight Glutenin Subunit in the Endoplasmic Reticulum

2008 ◽  
Vol 149 (1) ◽  
pp. 412-423 ◽  
Author(s):  
Alessio Lombardi ◽  
Alessandra Barbante ◽  
Pietro Della Cristina ◽  
Daniele Rosiello ◽  
Chiara Lara Castellazzi ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Endoplasmic Reticulum ◽  
Disulfide Bond ◽  
Bond Formation ◽  
Glutenin Subunit ◽  
Low Molecular Weight ◽  
Disulfide Bond Formation ◽  
Interchain Disulfide Bond
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