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Related Concept Videos

Rab Proteins01:14

Rab Proteins

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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.
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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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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.
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The Ras Gene02:38

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The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
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GPCRs Regulate Adenylyl Cylase Activity01:09

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Some GPCRs transmit signals through adenylyl cyclase (AC), a transmembrane enzyme. AC helps synthesize second messenger cyclic adenosine monophosphate (cAMP). AC catalyzes cyclization reaction and converts ATP to cAMP by releasing a pyrophosphate. The pyrophosphate is further hydrolyzed to phosphate by the enzyme pyrophosphatase, which drives cAMP synthesis to completion. However, cAMP is rapidly degraded to 5′ AMP by the enzymes phosphodiesterase (PDE), preventing overstimulation of...
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Cells migrating in response to external stimuli form lamellipodia, which are thin membrane protrusions supported by a mesh of linked, branched, or unbranched actin filaments. These actin filaments interact with myosin motor proteins, creating the dynamic actomyosin complex within the cytoskeleton. Contractility, or the ability to generate contractile stress, is inherent to the actomyosin complex. It helps cells detect the stiffness of the surrounding ECM and exert contractile force for...
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Related Experiment Video

Updated: Nov 20, 2025

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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Rab1-AMPylation by Legionella DrrA is allosterically activated by Rab1.

Jiqing Du1,2, Marie-Kristin von Wrisberg3, Burak Gulen1,2

  • 1Center for Integrated Protein Science Munich (CIPSM), Department of Chemistry, Technical University of Munich, Garching, 85748, Germany.

Nature Communications
|January 20, 2021
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Legionella bacteria use the DrrA protein to modify the host Rab1 protein via AMPylation. This study reveals a new binding site on DrrA that controls this modification, potentially preventing harmful host cell damage.

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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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Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods
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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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Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods
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Area of Science:

  • Microbiology
  • Structural Biology
  • Biochemistry

Background:

  • Legionella pneumophila infects host cells by forming a specialized vacuole.
  • The bacterial protein DrrA/SidM manipulates host vesicular trafficking regulator Rab1.
  • Rab1 undergoes adenosine monophosphate (AMP) modification, termed AMPylation, mediated by DrrA.

Purpose of the Study:

  • To investigate the molecular basis of the interaction between Rab1 and the DrrA AMPylation domain.
  • To understand how DrrA's activity is regulated during host cell infection.

Main Methods:

  • Utilized a chemical approach to stabilize low-affinity Rab:DrrA complexes site-specifically.
  • Determined the crystal structure of the Rab:DrrA complex.
  • Performed biochemical characterization of DrrA activity.

Main Results:

  • Identified a novel non-conventional Rab-binding site (NC-RBS) on DrrA.
  • Demonstrated that Rab1 binding to the NC-RBS allosterically stimulates DrrA's AMPylation activity.
  • The crystal structure provided insights into the Rab:DrrA interaction.

Conclusions:

  • Rab1 binding to the NC-RBS is crucial for DrrA-mediated AMPylation.
  • Allosteric control of DrrA may prevent potentially cytotoxic AMPylation in the host.
  • This regulation likely ensures efficient Legionella infection by controlling host cell processes.