The bundling of actin with polyethylene glycol 8000 in the presence and absence of gelsolin.
Identifieur interne : 003E05 ( Main/Curation ); précédent : 003E04; suivant : 003E06The bundling of actin with polyethylene glycol 8000 in the presence and absence of gelsolin.
Auteurs : J. Goverman [États-Unis] ; L A Schick ; J. NewmanSource :
- Biophysical journal [ 0006-3495 ] ; 1996.
Descripteurs français
- KwdFr :
- Actines (), Actines (physiologie), Animaux, Biophysique, Cytosol (), Cytosol (physiologie), Diffusion de rayonnements, Gelsoline (), Gelsoline (physiologie), Lapins, Lumière, Microscopie de contraste de phase, Phénomènes biophysiques, Polyéthylène glycols (), Structure moléculaire, Structures macromoléculaires, Techniques in vitro.
- MESH :
English descriptors
- KwdEn :
- Actins (chemistry), Actins (physiology), Animals, Biophysical Phenomena, Biophysics, Cytosol (chemistry), Cytosol (physiology), Gelsolin (chemistry), Gelsolin (physiology), In Vitro Techniques, Light, Macromolecular Substances, Microscopy, Phase-Contrast, Molecular Structure, Polyethylene Glycols (chemistry), Rabbits, Scattering, Radiation.
- MESH :
- chemical , chemistry : Actins, Gelsolin, Polyethylene Glycols.
- chemical , physiology : Actins, Gelsolin.
- chemistry : Cytosol.
- physiology : Cytosol.
- Animals, Biophysical Phenomena, Biophysics, In Vitro Techniques, Light, Macromolecular Substances, Microscopy, Phase-Contrast, Molecular Structure, Rabbits, Scattering, Radiation.
Abstract
Actin filament and bundle formation occur in the cytosol under conditions of very high total macromolecular concentration. In this study we have utilized the inert molecule polyethylene glycol 8000 (PEG) as a means of simulating crowded conditions in vitro. Column-purified Ca-actin was polymerized in the absence and presence of gelsolin (to regulate mean filament lengths between 50 and 5000 mers) and PEG (2-8%) using various concentrations of KCl and/or 2 mM divalent cations. Bundling was characterized by the scattered light intensity and mean diffusion coefficients obtained from dynamic light scattering, as well as by fluorescence and phase-contrast microscopy. The minimum concentration of KCl required for bundling decreases both with increasing concentration of PEG at a fixed mean filament length, and with decreasing filament length at a fixed concentration of PEG. In the absence of divalent cation, bundling is reversible on dilution, as determined by intensity levels, diffusion coefficients, and microscopy. However, with either 2 mM Mg2+ or Ca2+ added, bundling is irreversible under conditions of higher PEG concentrations or longer filaments, indicating that osmotic pressure effects cannot fully explain actin bundling with PEG. Weaker divalent cation-binding sites on actin as well as disulfide bonds appear to be involved in the irreversible bundling.
DOI: 10.1016/S0006-3495(96)79349-9
PubMed: 8874022
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pubmed:8874022Le document en format XML
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<term>Biophysical Phenomena</term>
<term>Biophysics</term>
<term>Cytosol (chemistry)</term>
<term>Cytosol (physiology)</term>
<term>Gelsolin (chemistry)</term>
<term>Gelsolin (physiology)</term>
<term>In Vitro Techniques</term>
<term>Light</term>
<term>Macromolecular Substances</term>
<term>Microscopy, Phase-Contrast</term>
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<term>Actines (physiologie)</term>
<term>Animaux</term>
<term>Biophysique</term>
<term>Cytosol ()</term>
<term>Cytosol (physiologie)</term>
<term>Diffusion de rayonnements</term>
<term>Gelsoline ()</term>
<term>Gelsoline (physiologie)</term>
<term>Lapins</term>
<term>Lumière</term>
<term>Microscopie de contraste de phase</term>
<term>Phénomènes biophysiques</term>
<term>Polyéthylène glycols ()</term>
<term>Structure moléculaire</term>
<term>Structures macromoléculaires</term>
<term>Techniques in vitro</term>
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<term>Gelsolin</term>
<term>Polyethylene Glycols</term>
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<term>Gelsolin</term>
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<term>Lumière</term>
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<term>Phénomènes biophysiques</term>
<term>Polyéthylène glycols</term>
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<front><div type="abstract" xml:lang="en">Actin filament and bundle formation occur in the cytosol under conditions of very high total macromolecular concentration. In this study we have utilized the inert molecule polyethylene glycol 8000 (PEG) as a means of simulating crowded conditions in vitro. Column-purified Ca-actin was polymerized in the absence and presence of gelsolin (to regulate mean filament lengths between 50 and 5000 mers) and PEG (2-8%) using various concentrations of KCl and/or 2 mM divalent cations. Bundling was characterized by the scattered light intensity and mean diffusion coefficients obtained from dynamic light scattering, as well as by fluorescence and phase-contrast microscopy. The minimum concentration of KCl required for bundling decreases both with increasing concentration of PEG at a fixed mean filament length, and with decreasing filament length at a fixed concentration of PEG. In the absence of divalent cation, bundling is reversible on dilution, as determined by intensity levels, diffusion coefficients, and microscopy. However, with either 2 mM Mg2+ or Ca2+ added, bundling is irreversible under conditions of higher PEG concentrations or longer filaments, indicating that osmotic pressure effects cannot fully explain actin bundling with PEG. Weaker divalent cation-binding sites on actin as well as disulfide bonds appear to be involved in the irreversible bundling.</div>
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