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Related Concept Videos

Tail-anchoring of Proteins in the ER Membrane01:45

Tail-anchoring of Proteins in the ER Membrane

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Tail-anchored, or TA, proteins are estimated to make up to 3-5% of membrane proteins found in the eukaryotic cell. Such proteins have a single transmembrane domain located approximately 30 amino acid residues upstream from the C-terminal end. As a result, the signal recognition particle (SRP) cannot guide a TA protein to the ER membrane for cotranslational insertion. Hence, they are integrated into the ER membrane post-translationally using their C-terminal end as the anchor. TA proteins...
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GPI Anchoring of Proteins in the ER Membrane01:29

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GPI-anchoring is a post-translational, reversible protein modification that is ubiquitous in eukaryotes. Such proteins are primarily present on the exoplasmic leaflet of the plasma membrane.
GPI-anchor structure
A sequence of 11 enzymatic reactions results in the synthesis of the complete GPI anchor consisting of a hydrophobic and a hydrophilic portion. The hydrophobic portion comprises phosphatidylinositol, while the hydrophilic part comprises polar groups like phosphoethanolamine,...
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Protein Translocation Machinery on the ER Membrane01:28

Protein Translocation Machinery on the ER Membrane

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The translocon complex situated on the ER membrane is the main gateway for the protein secretory pathway. It facilitates the transport of nascent peptides into the ER lumen and their insertion into the ER membrane.
Sec61 protein conducting channel
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Insertion of Single-pass Transmembrane Proteins in the RER01:26

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Integral membrane proteins are proteins adhered to the lipid bilayer of a cell organelle or membrane. They can be of two types: transmembrane integral proteins that span the lipid bilayer and monotopic proteins that are attached to either side of the membrane but do not pass through it.
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Protein Transport into the Inner Mitochondrial Membrane01:34

Protein Transport into the Inner Mitochondrial Membrane

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Nuclear encoded mitochondrial precursors are imported to the inner membrane in a multistep process involving two separate translocons, TIM22 and TIM23. TIM23 is a cation-selective pore that remains closed by the N terminal segment of the protein. Negative charges on the TIM23 act as a receptor for the incoming precursor, pulling the positively charged matrix-targeting sequence for peptide insertion and translocation.
Transport of mitochondrial precursors across the TIM23 channel is driven by...
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Cotranslational Protein Translocation01:20

Cotranslational Protein Translocation

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Translocation of proteins across membranes is an ancient process that occurs even in bacteria and archaebacteria. In fact, the components of the translocation machinery are still conserved between prokaryotes and eukaryotes.
Sec61 channel partners for cotranslational translocation
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Related Experiment Video

Updated: Feb 23, 2026

Reconstitution of Msp1 Extraction Activity with Fully Purified Components
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Tail-Anchored Protein Insertion by a Single Get1/2 Heterodimer.

Benjamin E Zalisko1, Charlene Chan2, Vladimir Denic2

  • 1Department of Biochemistry and Molecular Biology, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.

Cell Reports
|September 7, 2017
PubMed
Summary

A single Get1/2 heterodimer is sufficient for inserting tail-anchored proteins into the endoplasmic reticulum membrane. This complex binds Get3 asymmetrically, clarifying protein insertion mechanisms.

Keywords:
GET pathwayGet1/2 transmembrane complexbiogenesismembrane protein insertionmolecular mechanismproteoliposomessingle-molecule FRETtail-anchored protein

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Transmembrane Domain Oligomerization Propensity determined by ToxR Assay
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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Protein Trafficking

Background:

  • Tail-anchored (TA) proteins insert into the endoplasmic reticulum membrane via the Get1/2 complex and Get3 chaperone.
  • The precise architecture and mechanism of the Get1/2 complex in TA protein insertion remain poorly understood.

Purpose of the Study:

  • To determine the oligomeric state and architecture of the full-length Get1/2 complex.
  • To elucidate the coordination mechanism between Get1/2 and Get3 during TA protein insertion.

Main Methods:

  • Reconstitution of the Get1/2 complex in lipid bilayers.
  • Single-molecule and bulk fluorescence measurements.
  • Quantitative in vitro insertion assays.

Main Results:

  • A single Get1/2 heterodimer is sufficient for TA protein insertion.
  • The cytosolic regions of Get1 and Get2 bind Get3 asymmetrically.
  • Defined the stoichiometry of the Get1/2-Get3 interaction.

Conclusions:

  • Established a simplified model for Get1/2 and Get3 coordinating TA protein insertion.
  • Provided mechanistic insight into the Get1/2 complex's role in membrane protein biogenesis.