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Related Concept Videos

ABC Transporters: Exporter01:31

ABC Transporters: Exporter

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ATP-binding cassette or ABC transporter is the largest superfamily of integral membrane proteins. The transporters have transmembrane-binding domains (TMDs) and nucleotide-binding domains (NBDs). The TMDs are specific to their substrates, whereas the NBDs are similar to engines that complete ATP hydrolysis to complete the substrate transport. They can be full transporters consisting of two TMDs and NBDs, half transporters with one TMD and NBD, while some encoded with a single TMD or NBD are...
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ABC Transporters: Importer01:27

ABC Transporters: Importer

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ATP-binding cassette or ABC transporters are a class of ATP-driven pumps that hydrolyze ATP to move solutes across the membrane. They can be grouped into importers and exporters. While exporters are present in all domains of life, importers exist only in bacteria and some plants.
In bacteria, based on the number of transmembrane helices and the chemical nature of their substrates, the ABC importers can be divided into three types:
3.6K
Carrier-Mediated Transport01:06

Carrier-Mediated Transport

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Carrier-mediated transport is a pivotal process in drug absorption, particularly for lipid-insoluble drugs, and encompasses facilitated diffusion and active transport. Facilitated diffusion allows drugs to move along their concentration gradient without energy expenditure, while active transport utilizes ATP to drive drug movement against this gradient.
Active transport involves two types of membrane-spanning transporters: uptake and efflux. Uptake transporters are expressed in the small...
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Drug Absorption Mechanism: Carrier-Mediated Membrane Transport01:19

Drug Absorption Mechanism: Carrier-Mediated Membrane Transport

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Certain large, lipid-insoluble drug molecules that resemble amino acids, peptides, or glucose, require specialized carrier proteins to facilitate their diffusion across cell membranes. This transport can occur through either facilitated diffusion, which does not require energy input, or active transport, which does require energy input.
Facilitated diffusion is a passive process that utilizes human Solute Carrier (SLC) transporters. These transporters bind to the drug, undergo structural...
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Nuclear Export01:42

Nuclear Export

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The nucleus restricts several proteins within and allows others to pass. The restricted proteins possess a nuclear retention sequence or NRS, anchoring them to the nuclear lamins and preventing their transport to the cytosol. The non-restricted proteins, after their synthesis, are transported to their site of action, such as the cytosol or other organelles, with the help of nuclear export signals or NES.
NES are of three types- the canonical 10-residue long leucine-rich signal and other...
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Mechanisms of Drug Absorption: Paracellular, Transcellular, and Vesicular Transport01:23

Mechanisms of Drug Absorption: Paracellular, Transcellular, and Vesicular Transport

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Drugs need to permeate cell membranes to reach their target sites after administration. Orally administered drugs must transcend intestinal epithelial membrane barriers to infiltrate the systemic circulation. Drugs with a molecular weight of less than 500 Daltons diffuse through gaps between neighboring cells, called paracellular pathways.
However, most drugs use the transcellular route, traversing directly through the cell membranes via two mechanisms: passive and active transport. Passive...
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Related Experiment Video

Updated: Mar 7, 2026

Cholesterol Efflux Assay
07:54

Cholesterol Efflux Assay

Published on: March 6, 2012

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ABC-Transporter Mediated Sterol Export from Cells Using Radiolabeled Sterols.

Alryel Yang1, Ingrid C Gelissen2

  • 1Faculty of Pharmacy, The University of Sydney, Pharmacy Bank Building A15, Camperdown, Sydney, NSW, 2006, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|February 17, 2017
PubMed
Summary

This study details methods for measuring cellular cholesterol export, a key step in reverse cholesterol transport. These techniques use radiolabeled sterols to assess ATP-binding cassette (ABC) transporter function in controlling cholesterol levels.

Keywords:
ABC-transporterABCA1ABCA7ABCG1ABCG4Cholesterol exportReverse cholesterol transport

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High-throughput Nitrobenzoxadiazole-labeled Cholesterol Efflux Assay
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Related Experiment Videos

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Cholesterol Efflux Assay
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Author Spotlight: A Bicelle Crystallization Setup for ABC Transporter Membrane Proteins to Advance Drug Development
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High-throughput Nitrobenzoxadiazole-labeled Cholesterol Efflux Assay
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High-throughput Nitrobenzoxadiazole-labeled Cholesterol Efflux Assay

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Area of Science:

  • Cellular biology
  • Biochemistry
  • Molecular medicine

Background:

  • Cellular cholesterol homeostasis is critical for preventing diseases like atherosclerosis.
  • Cholesterol export is the initial and vital step in reverse cholesterol transport.
  • Dysfunctional cholesterol transport leads to cellular lipid accumulation.

Purpose of the Study:

  • To describe methodologies for quantifying sterol export mediated by ATP-binding cassette (ABC) transporters.
  • To provide tools for studying the role of specific ABC transporters (ABCA1, ABCA7, ABCG1, ABCG4) in cholesterol metabolism.
  • To enable the assessment of transporter function in both endogenous and overexpression systems.

Main Methods:

  • Utilizing radiolabeled sterols to trace and quantify cholesterol movement.
  • Employing techniques to measure sterol export from cells to extracellular acceptors.
  • Comparing sterol export in cells with naturally expressed transporters versus those with overexpressed transporters.

Main Results:

  • Established protocols for measuring ABC-transporter-mediated sterol export.
  • Demonstrated the applicability of radiolabeled sterol methods in diverse cellular models.
  • Provided a framework for investigating the functional impact of ABC transporters on cholesterol efflux.

Conclusions:

  • Radiolabeled sterol assays are effective for studying ABC transporter-mediated cholesterol export.
  • These methods are crucial for understanding reverse cholesterol transport and its role in health and disease.
  • The described techniques facilitate research into therapeutic strategies targeting cholesterol metabolism.