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Related Concept Videos

Export of Misfolded Proteins out of the ER01:32

Export of Misfolded Proteins out of the ER

After folding, the ER assesses the quality of secretory and membrane proteins. The correctly folded proteins are cleared by the calnexin cycle for transport to their final destination, while misfolded proteins are held back in the ER lumen. The ER chaperones attempt to unfold and refold the misfolded proteins but sometimes fail to achieve the correct native conformation. Such terminally misfolded proteins are then exported to the cytosol by ER-associated degradation or ERAD pathway for...
Amyloid Fibrils03:03

Amyloid Fibrils

Amyloid fibrils are aggregates of misfolded proteins.  Under most circumstances, misfolded proteins are either refolded by chaperone proteins or degraded by the proteasome. However, in the case of a mutation or a disease, these proteins can accumulate to form large clusters and often further assemble to form elongated fibers, called fibrils. 
Amyloid deposits were observed as early as 1639 in the liver and the spleen.   In 1854, Rudolph Virchow performed iodine staining, normally used to...
Amyloid Fibrils03:03

Amyloid Fibrils

Amyloid fibrils are aggregates of misfolded proteins.  Under most circumstances, misfolded proteins are either refolded by chaperone proteins or degraded by the proteasome. However, in the case of a mutation or a disease, these proteins can accumulate to form large clusters and often further assemble to form elongated fibers, called fibrils. 
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The Unfolded Protein Response01:37

The Unfolded Protein Response

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...
Molecular Chaperones and Protein Folding03:00

Molecular Chaperones and Protein Folding

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...
Bacterial Protein Maturation01:26

Bacterial Protein Maturation

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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Related Experiment Video

Updated: Jun 4, 2026

Protein Misfolding Cyclic Amplification of Prions
10:12

Protein Misfolding Cyclic Amplification of Prions

Published on: November 7, 2012

Highly efficient protein misfolding cyclic amplification.

Nuria Gonzalez-Montalban1, Natallia Makarava, Valeriy G Ostapchenko

  • 1Center for Biomedical Engineering and Technology, University of Maryland, Baltimore, Maryland, United States of America.

Plos Pathogens
|February 25, 2011
PubMed
Summary
This summary is machine-generated.

Introducing Teflon beads into protein misfolding cyclic amplification (PMCA) dramatically boosts prion replication efficiency. This modified PMCA (PMCAb) enhances prion detection sensitivity and yield for research and diagnostics.

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Last Updated: Jun 4, 2026

Protein Misfolding Cyclic Amplification of Prions
10:12

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Published on: November 7, 2012

Assays for the Degradation of Misfolded Proteins in Cells
10:56

Assays for the Degradation of Misfolded Proteins in Cells

Published on: August 28, 2016

Purification of Hsp104, a Protein Disaggregase
07:17

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Published on: September 30, 2011

Area of Science:

  • Neuroscience
  • Biochemistry
  • Molecular Biology

Background:

  • Protein misfolding cyclic amplification (PMCA) is a key method for in vitro prion replication.
  • Low conversion efficiency of normal prion protein (PrP(C)) to misfolded form (PrP(Sc)) limits PMCA applications.
  • This inefficiency suggested potential limitations in substrate availability or cofactor concentrations.

Purpose of the Study:

  • To improve the efficiency, rate, and robustness of prion conversion in PMCA.
  • To enhance the sensitivity of prion detection assays.
  • To investigate the underlying reasons for previously observed low conversion efficiencies.

Main Methods:

  • Modification of the PMCA protocol by incorporating Teflon beads, creating the PMCAb method.
  • Quantification of PrP(C) to PrP(Sc) conversion rates and amplification factors.
  • Assessment of prion detection sensitivity using serial dilutions of infected brain material.

Main Results:

  • PMCAb increased PrP(C) to PrP(Sc) conversion from ~10% to up to 100%.
  • A single 24-hour PMCAb round achieved 600-700-fold amplification of PrP(Sc).
  • Prion detection sensitivity increased by 2-3 orders of magnitude in 24 hours; a 10^12 dilution was detectable in 72 hours.
  • Improved efficiency was observed for both hamster 263K and synthetic SSLOW prion strains.
  • Enhanced amplification did not compromise prion replication specificity.

Conclusions:

  • The addition of Teflon beads (PMCAb) significantly enhances prion replication and detection.
  • Low conversion efficiency is not due to limited susceptible PrP(C) or essential cofactors.
  • PMCAb offers a robust platform for ultrasensitive prion detection and in vitro PrP(Sc) production.