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G Protein-coupled Receptors01:15

G Protein-coupled Receptors

16.2K
G Protein-Coupled Receptors or GPCRs are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to sensory stimuli such as light, odors, hormones, cytokines, or neurotransmitters.
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
16.2K
Transducer Mechanism: G Protein–Coupled Receptors01:30

Transducer Mechanism: G Protein–Coupled Receptors

3.7K
G Protein–Coupled Receptors (GPCRs) are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to various stimuli. GPCRs regulate critical physiological pathways and are excellent drug targets for treating diseases such as diabetes, cancer, obesity, depression, or Alzheimer's. Nearly 35% of approved drugs implement their therapeutic effects by selectively interacting with specific GPCRs.
GPCRs are also called heptahelical,...
3.7K
Protein Networks02:26

Protein Networks

4.4K
An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
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Proteomics01:33

Proteomics

9.2K
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
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G-protein Coupled Receptors01:21

G-protein Coupled Receptors

131.2K
G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.
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Related Experiment Videos

Seven perspectives on GPCR H/D-exchange proteomics methods.

Xi Zhang1

  • 1Independent Researcher, Montreal, QC, H2Y 1H3, Canada.

F1000Research
|May 23, 2017
PubMed
Summary
This summary is machine-generated.

This article clarifies technical nuances for visualizing human G protein-coupled receptor (GPCR) dynamic structures using hydrogen deuterium exchange (HDX) proteomics. It guides scientists to overcome challenges and reproduce high-quality GPCR HDX structural analysis.

Keywords:
GPCRH/D-exchangedetergentslipidsmembrane proteinsstructural proteomics

Related Experiment Videos

Area of Science:

  • Structural Biology
  • Proteomics
  • Biochemistry

Background:

  • Growing interest in visualizing dynamic structures of human G protein-coupled receptors (GPCRs).
  • Hydrogen-deuterium exchange (HDX) proteomics offers a powerful bottom-up approach for structural studies.
  • Challenges exist in applying HDX proteomics to membrane proteins like GPCRs due to technical nuances.

Purpose of the Study:

  • To clarify critical technical nuances and logical thinking behind the GPCR HDX proteomics method.
  • To assist scientists in overcoming cross-discipline pitfalls for high-quality protocol reproduction.
  • To emphasize considerations for membrane protein stability and compatibility throughout the HDX pipeline.

Main Methods:

  • Systematic consideration of membrane protein stability, detergent buffers, and chromatography settings.
  • Viewing bottom-up HDX as two steps: (I) protein HDX labeling in native buffers, and (II) peptide-centric analysis.
  • Utilizing bottom-up MS/MS for peptide matrix construction and HDX-MS for label localization and quantification.
  • Application of a detergent-low-TCEP digestion method for improved GPCR digestion.

Main Results:

  • Achieved 89% structural coverage of GPCRs in 2010 using rigorous HDX methods.
  • Highlighted the impact of metal ions, zero-detergent shock, and freeze-thaws on HDX result rigor.
  • Demonstrated a structured approach where experimental conditions prioritize protein structure.

Conclusions:

  • GPCR HDX proteomics is a robust structural approach requiring careful optimization.
  • Understanding and addressing technical nuances are crucial for accurate and reproducible structural insights.
  • The proposed two-step method and specific digestion technique facilitate successful GPCR HDX analysis.