Protein Crowding
Protein Crowding - Related Videos
10:23
Crowding
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2023
Source: Laboratory of Jonathan Flombaum—Johns Hopkins University Human vision depends on light-sensitive neurons that are arranged in the back of the eye on a tissue called the retina. The neurons, called the rods and cones because of their shapes, are not uniformly distributed on the retina. Instead, there is a region in the center of the retina called the macula where cones are densely packed, and especially so in a central sub-region of the macula called the fovea. Outside the fovea there...
09:14
Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding
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Cited by 14 •
2016
We present a protocol to obtain cell-derived matrices rich in extracellular matrix proteins, using macromolecular crowders (MMC). In addition, we present a protocol which incorporates MMC in 3D organotypic skin co-culture generation, which reduces culture time while maintaining maturity of construct.
08:46
Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
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Cited by 34 •
2015
A streamlined approach to screening for the expression of recombinant membrane proteins in Escherichia coli based on fusion to green fluorescent protein is presented.
08:20
In Vitro Model of Human Cutaneous Hypertrophic Scarring using Macromolecular Crowding
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Cited by 8 •
2020
This protocol describes the use of macromolecular crowding to create an in vitro human hypertrophic scar tissue model that resembles in vivo conditions. When cultivated in a crowded macromolecular environment, human skin fibroblasts exhibit phenotypes, biochemistry, physiology, and functional characteristics resembling scar tissue.
10:43
Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes
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2022
Here, a protocol to perform and analyze the binding, mobility, and assembly of single molecules on artificial crowded lipid membranes using single-molecule total internal reflection fluorescence (smTIRF) microscopy is presented.
FAQs
Excluded volume acts by making part of the surrounding space unavailable to other molecules. As the local protein concentration rises, potential movement pathways become more constrained, so diffusion can change. This spatial restriction does not predict every outcome by itself; the resulting behavior also depends on protein size, shape, and nonspecific interactions.
Protein crowding can either promote or oppose molecular association. Reduced available space may bring molecules into closer effective proximity, while nonspecific interactions can determine whether that proximity leads to association. Solvent conditions and the physical characteristics of the proteins further influence the balance, so crowding is not universally association-enhancing.
Effects observed in crowded biological environments may differ from measurements made in dilute laboratory solutions. Protein crowding can change how folding, enzyme activity, and aggregation are interpreted because these processes occur within restricted space and under interaction conditions that dilute measurements may not reproduce. This comparison helps relate laboratory findings to intracellular behavior.
Researchers can compare the same biomolecular behavior under crowded and dilute conditions, then examine changes in diffusion, association, folding, enzyme activity, or aggregation. Interpreting those differences requires attention to protein size and shape, solvent conditions, and nonspecific interactions, because these factors can shift both the direction and magnitude of the observed effect.
Protein size and shape influence how much available space is effectively occupied and how other molecules encounter that protein. Consequently, two crowded environments with similar overall concentration may not produce identical outcomes. Considering these physical differences is important when interpreting changes in diffusion, molecular association, folding, or aggregation.
Protein crowding provides context for several processes studied in biology, including protein folding, enzyme activity, aggregation, molecular transport, and the organization of complex intracellular environments. Its relevance comes from showing why these behaviors may depend on the surrounding molecular density and interactions, rather than matching results obtained in simplified dilute solutions.