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Related Concept Videos

Molecular Chaperones and Protein Folding03:00

Molecular Chaperones and Protein Folding

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The native conformation of a protein is formed by interactions between the side chains of its constituent amino acids. When the amino acids cannot form these interactions, the protein cannot fold by itself and needs chaperones. Notably, chaperones do not relay any additional information required for the folding of polypeptides; the native conformation of a protein is determined solely by its amino acid sequence. Chaperones catalyze protein folding without being a part of the folded protein.
The...
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Protein Folding01:25

Protein Folding

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Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation, critical to its biological function. Interactions between its constituent amino acids guide protein folding, and hence the protein structure is primarily dependent on its amino acid sequence.
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
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Bacterial Protein Maturation01:26

Bacterial Protein Maturation

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Bacterial protein maturation is a tightly regulated process that ensures newly synthesized polypeptides achieve correct functional conformations. This maturation involves a series of modifications, folding events, and quality control steps, often assisted by specialized chaperone proteins.N-Terminal ModificationsThe maturation of bacterial polypeptides begins cotranslationally as the polypeptide exits the ribosome. The first amino acid, N-formylmethionine (fMet), is typically modified at the...
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Protein Folding Quality Check in the RER01:29

Protein Folding Quality Check in the RER

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ER is the primary site for the maturation and folding of soluble and transmembrane secretory proteins. The calnexin cycle is a specific chaperone system that folds and assesses the confirmation of N-glycosylated proteins before they can exit the ER lumen. The primary players of this quality check pipeline are the lectins, ER-resident chaperones, and a glucosyl transferase enzyme. In case the calnexin system in the lumen fails to salvage a misfolded protein, it is transported to the cytoplasm...
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Post-translational Translocation of Proteins to the RER01:27

Post-translational Translocation of Proteins to the RER

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A sizable fraction of proteins destined for ER are first synthesized in the cell cytosol and then transported across the ER membrane–a process called post-translational translocation. Similar to cotranslationally translocated proteins, these proteins also use the Sec translocon complex to enter the ER lumen.
Targeting proteins to the ER
Hsp40 and Hsp70 chaperone molecules bind the translated proteins in the cytosol to prevent their folding. The chaperone binding helps to keep the signal...
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The Unfolded Protein Response01:37

The Unfolded Protein Response

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The ER is the hub of protein synthesis in a cell. It has robust systems to quality control protein folding and also for degradation of terminally misfolded proteins. Under normal conditions, a small proportion of misfolded proteins that cannot be salvaged need to be transported to the cytoplasm by the ER-associated degradation or ERAD pathways. However, if the ERAD cannot handle the misfolded proteins, the cell activates the unfolded protein response or UPR to adjust the protein folding...
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Related Experiment Video

Updated: Sep 14, 2025

Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides
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Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides

Published on: June 19, 2012

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Arrest Peptide Profiling resolves co-translational folding pathways and chaperone interactions in vivo.

Xiuqi Chen1,2,3, Vincent J Hilser2, Christian M Kaiser4,5

  • 1CMDB Graduate Program, Johns Hopkins University, Baltimore, MD, USA.

Nature Communications
|July 24, 2025
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Summary

We developed Arrest Peptide Profiling (AP Profiling) to study protein folding as it happens during translation. This method reveals how protein structure and molecular chaperones influence folding pathways in live cells.

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In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells
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Analysis of Protein Folding, Transport, and Degradation in Living Cells by Radioactive Pulse Chase
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Analysis of Protein Folding, Transport, and Degradation in Living Cells by Radioactive Pulse Chase

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Related Experiment Videos

Last Updated: Sep 14, 2025

Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides
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Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides

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In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells
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Analysis of Protein Folding, Transport, and Degradation in Living Cells by Radioactive Pulse Chase
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Analysis of Protein Folding, Transport, and Degradation in Living Cells by Radioactive Pulse Chase

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Area of Science:

  • Molecular biology
  • Biochemistry
  • Cell biology

Background:

  • Cytosolic proteins initiate folding co-translationally as they emerge from ribosomes.
  • This early folding is critical for protein structure and function, guided by molecular chaperones.
  • Detecting and understanding co-translational folding in real-time remains a significant challenge.

Purpose of the Study:

  • To quantitatively analyze co-translational protein folding dynamics in live cells.
  • To elucidate how protein topology influences folding pathways.
  • To investigate the role of nascent chain-binding chaperones in protein folding.

Main Methods:

  • Development of a high-throughput method: Arrest Peptide Profiling (AP Profiling).
  • Integration of AP Profiling with single-molecule experiments.
  • Analysis of GTPase domains to understand folding pathway determinants.

Main Results:

  • AP Profiling successfully defined co-translational folding for GTPase domains.
  • Protein topology was shown to shape folding pathways.
  • Ablation of specific chaperones revealed distinct, localized folding changes, explaining chaperone functional redundancy.

Conclusions:

  • This study provides unprecedented insight into cellular folding pathways of complex proteins.
  • AP Profiling enables systematic studies of nascent protein folding with high resolution and throughput.
  • The findings advance our understanding of protein biogenesis and quality control in cells.