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Related Concept Videos

Rab Proteins01:14

Rab Proteins

4.1K
Rab proteins constitute the largest family of monomeric GTPases, of which 70 members are present in humans. Rab proteins and their effectors regulate consecutive stages of vesicle transport such as vesicle transport, docking, and fusion to the correct recipient membrane.
Rab proteins switch between a cytosolic, GDP-bound inactive state and a membrane-anchored, GTP-bound active state. By themselves, Rabs show slow rates of GDP/GTP exchange and GTP hydrolysis. Thus, Rab proteins are considered...
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Rab Cascades01:25

Rab Cascades

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Rab GTPases act in a regulated cascade during membrane fusion, helping the lipid bilayers mix. The Rab family of proteins are active when bound to GTP, and inactive when bound to GDP. Hence, they act as guanine nucleotide-dependent molecular switches. Rab-GTP recognizes and binds to long or short-range tethering proteins to capture the target vesicle. These tethers coordinate with SNAREs on the vesicle and the target membrane to assemble the trans SNARE complex that locks the mixing bilayers.
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Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

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Ras and Rho are small monomeric GTPases that act downstream of receptor tyrosine kinase (RTK) and regulate various cellular processes. These GTPases switch between active and inactive states by binding to guanine nucleotides.
Three regulatory proteins control their activity:
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Coat Assembly and GTPases01:33

Coat Assembly and GTPases

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Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
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Directing Proteins to the Rough Endoplasmic Reticulum01:34

Directing Proteins to the Rough Endoplasmic Reticulum

7.4K
The organelle-specific signaling sequences direct proteins synthesized in the cytosol to their final destination like ER, mitochondria, peroxisomes, etc. Some of the proteins directed to ER are then trafficked via vesicles to other organelles within the cell or the extracellular environment through the Golgi complex. For example, the rough ER synthesizes soluble proteins for transportation to the lysosomes or secretion out of the cell. It can also synthesize transmembrane proteins that can...
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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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Related Experiment Video

Updated: Sep 3, 2025

Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein GST-RhoAG17A from Epithelial Cell Lysates
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Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein GST-RhoAG17A from Epithelial Cell Lysates

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Rep15 interacts with several Rab GTPases and has a distinct fold for a Rab effector.

Amrita Rai1, Anurag K Singh2, Nathalie Bleimling3

  • 1Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany. amrita.rai@mpi-dortmund.mpg.de.

Nature Communications
|July 23, 2022
PubMed
Summary
This summary is machine-generated.

Researchers characterized Rep15, a Rab15 effector protein, identifying new interactions with Rab3 paralogs and Rab34. This study reveals Rep15

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Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
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Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

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Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells
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Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells

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Related Experiment Videos

Last Updated: Sep 3, 2025

Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein GST-RhoAG17A from Epithelial Cell Lysates
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Affinity Precipitation of Active Rho-GEFs Using a GST-tagged Mutant Rho Protein GST-RhoAG17A from Epithelial Cell Lysates

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Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
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Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells
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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Structural Biology

Background:

  • Rab proteins regulate downstream signaling through interactions with effector proteins.
  • Rep15 is a known Rab15 effector involved in receptor recycling but is poorly understood.
  • Rab proteins are key regulators of intracellular membrane trafficking.

Purpose of the Study:

  • To characterize the interaction between Rep15 and Rab proteins.
  • To identify novel interacting partners of Rep15.
  • To elucidate the structural basis and functional significance of Rep15-Rab interactions.

Main Methods:

  • Yeast two-hybrid assay for identifying protein interactions.
  • Biochemical validation of identified interactions.
  • X-ray crystallography to determine complex structures.
  • Structure-based mutagenesis to analyze interaction specificity.
  • Cellular assays involving Rep15 depletion in glioblastoma cells.

Main Results:

  • Rep15 interacts with Rab15, Rab3 paralogs (Rab3B, Rab3C), and Rab34.
  • Crystal structures reveal Rep15's unique globular conformation and interaction interfaces.
  • Mutagenesis studies explain the specificity of Rep15-Rab interactions.
  • Rep15 depletion inhibits glioblastoma cell proliferation, migration, and receptor recycling.

Conclusions:

  • Rep15 is a multi-Rab effector with a distinct structural profile.
  • The Rep15:Rab interaction mechanism is elucidated, highlighting specificity determinants.
  • Rep15 plays a crucial role in glioblastoma cell functions, suggesting potential implications in cancer.