Detergent-resistant Membranes
Detergent-resistant Membranes - Related Videos
10:43
Expression, Detergent Solubilization, and Purification of a Membrane Transporter, the MexB Multidrug Resistance Protein
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Cited by 5 •
2010
In this protocol we demonstrate the expression, solubilization, and purification of a recombinantly expressed membrane protein, MexB, as a soluble protein detergent complex. MexB is a multidrug resistance membrane transporter from the opportunistic bacterial pathogen Pseudomonas aeruginosa.
01:18
Detergent Purification of Membrane Proteins
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2023
Detergents are used to purify the integral proteins of the membrane. The hydrophobic portion of the detergent can replace membrane phospholipids while solubilizing the membrane proteins. When detergent monomers reach a specific concentration in a solution called critical micelle concentration (CMC), they form micelles. Above CMC, the concentration of the detergent monomers remains in equilibrium with the micelle. The number of detergent monomers present in the CMC varies for each detergent, and...
10:09
Detection of Detergent-sensitive Interactions Between Membrane Proteins
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Cited by 2 •
2018
We describe a protocol for detection of detergent-sensitive interactions between membrane proteins using binding of the sorting receptor, sortilin, to the first luminal loop of the glucose transporter protein, GLUT4, as an example.
03:05
Fluorescence-Detection Size-Exclusion Chromatography: A Technique to Identify the Integrity of Fluorescent Membrane Proteins upon Detergent Solubilization
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2025
In this video, we demonstrate a fluorescence-detection size-exclusion chromatography technique to screen for green fluorescent protein (GFP)-fused membrane protein stability in Escherichia coli following detergent solubilization. Proteins exhibiting a symmetrical peak in the size-exclusion profile with little or no free GFP indicate purified proteins with minimum degradation and aggregation, which can subsequently be selected for structure-function analysis.
01:23
Synergistic Antimicrobial Effect of Membrane-Targeting Compounds on Antibiotic-Resistant Bacteria
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2026
Take suspensions of an antibiotic-resistant strain of a pathogenic bacterium.Treat one suspension with a short, membrane-disrupting antibiotic compound and the other with both the short compound and a longer, membrane-intercalating compound.Incubate both suspensions with agitation. These compounds mimic the lipid bilayer structure, enabling their spontaneous insertion into bacterial membranes.The short compound spans only part of the membrane’s thickness, creating a structural mismatch. This...
FAQs
Their resistance arises from tightly packed lipid interactions within ordered membrane regions. Cholesterol and sphingolipids contribute to this packing, allowing the fraction to remain relatively insoluble when exposed to selected nonionic detergents. Temperature also matters: extraction at low temperatures favors preservation of these ordered interactions, making detergent conditions an important part of the experimental interpretation.
Low temperatures are especially important because they support the ordered lipid interactions associated with detergent resistance. Changing the extraction temperature can therefore alter which membrane material remains insoluble and how strongly it is recovered. Researchers must consider this condition when comparing fractions, since resistance reflects both membrane composition and the physical conditions used during detergent treatment.
Detergent extraction creates an experimental fraction under conditions that may reorganize or selectively preserve membrane components. Consequently, insolubility is evidence of a particular biochemical behavior, not direct proof that an identical domain exists unchanged inside a living cell. This distinction helps researchers use the method as a model for membrane organization without treating it as a complete description of cellular membranes.
Researchers expose cell-membrane material to a suitable nonionic detergent, commonly under low-temperature conditions, and separate the relatively insoluble material from detergent-soluble components. They then isolate and analyze the resulting fractions to examine their lipid and protein content. The workflow provides a practical way to compare membrane components according to their behavior during extraction.
Fraction analysis can show whether particular membrane proteins partition with the ordered, detergent-resistant material or remain in detergent-soluble components. These patterns help investigators examine how proteins associate with membrane regions and how that organization may relate to signal transduction or vesicular trafficking. Interpretation remains comparative, because fractionation behavior alone does not establish a protein's location in intact cells.
The approach is useful when researchers need an experimentally tractable model of membrane organization in biology. By isolating and analyzing resistant fractions, they can investigate membrane rafts, protein partitioning, signal transduction, and vesicular trafficking. These applications connect lipid organization with cellular function, while the method's limitations encourage complementary interpretation rather than treating the fractions as exact replicas of living-cell domains.