Study of the translational diffusion of macromolecules in beads of gel chromatography by the FRAP method
We measured the translational diffusion of fractions of dextrans labelled with fluorescein isothiocyanate, in Sephadex gel beads permeated by aqueous solutions of these molecules. The molecular weights of these fractions were between 5400 and 200 000 and measurements of their diffusion coefficients...
Gespeichert in:
Veröffentlicht in: | Biophysical chemistry 1988-09, Vol.31 (3), p.247-258 |
---|---|
Hauptverfasser: | , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 258 |
---|---|
container_issue | 3 |
container_start_page | 247 |
container_title | Biophysical chemistry |
container_volume | 31 |
creator | Poitevin, Eric Wahl, Philippe |
description | We measured the translational diffusion of fractions of dextrans labelled with fluorescein isothiocyanate, in Sephadex gel beads permeated by aqueous solutions of these molecules. The molecular weights of these fractions were between 5400 and 200 000 and measurements of their diffusion coefficients inside a gel bead (
D) and in the free solution (
D
0), were performed using the fluorescence recovery after photobleaching method (FRAP). We also determined the coefficient of partitioning (
K
av) of these fractions between the gel and the free solvent, with a new microfluorimetric method. We found that, for Sephadex G-50, G-75, G-100, G-150 and G-200 gels,
K
av varied with the Stokes radius (
r
s) of the dextran molecules, in agreement with the formula of Laurent and Killander (J. Chromatogr. 14 (1964) 317). For Sephadex G-100, G-150 and G-200 gels,
D/
D
0 varied with r
s according to the theory of Ogston et al. (Proc. R. Soc. Lond. 333 (1973) 297). In addition, these theories predict a relation linking
D/
D
0 to
K
av which was wen verified. Our work is the first systematic study of the translational diffusion of macromolecules in a chromatography gel. These measurements should allow a better evaluation of the factors which influence the resolution in exclusion chromatography. In addition, the diffusion of macromolecules in gels may provide models for the diffusion of these molecules in the cytoplasm of living cells and in connective biological tissues. |
doi_str_mv | 10.1016/0301-4622(88)80030-9 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_78705207</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>0301462288800309</els_id><sourcerecordid>15132098</sourcerecordid><originalsourceid>FETCH-LOGICAL-c417t-395e7b61cbcb3f070611a139036bd37cf7416c37d7484c8840dc8652114ab9593</originalsourceid><addsrcrecordid>eNqFkd-LFSEUxyWK7bb1HxT4EFEPUzo66rwEy9JWsFD041kcPbNjOONNneD-9zl7L_exfNHD9-Ph8DkIPafkLSVUvCOM0IaLtn2t1BtFatn0D9COKska3hLyEO3OyGP0JOdfpJ4KXqCLlgvBe7lD0_eyugOOIy4T4JLMkoMpPi4mYOfHcc31vcWzsSnOMYBdA2TsFzyAcXmL7iBgO9XUlHiXzH464OFw3-_m29VXPEOZonuKHo0mZHh2ui_Rz5sPP64_NbdfPn6-vrptLKeyNKzvQA6C2sEObCSSCEoNZT1hYnBM2lFyKiyTTnLFrVKcOKtE11LKzdB3PbtEr4599yn-XiEXPftsIQSzQFyzlkqSriXyvyDtKGtJryrIj2AVkHOCUe-Tn006aEr0tgm9adabZq2Uvt-E3gZ5ceq_DjO486eT-pq_POUmWxPGqt76fMaElJT0bcXeHzGo0v54SDpbD4sF5xPYol30_57jL8Jzo3Q</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>15132098</pqid></control><display><type>article</type><title>Study of the translational diffusion of macromolecules in beads of gel chromatography by the FRAP method</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals Complete</source><creator>Poitevin, Eric ; Wahl, Philippe</creator><creatorcontrib>Poitevin, Eric ; Wahl, Philippe</creatorcontrib><description>We measured the translational diffusion of fractions of dextrans labelled with fluorescein isothiocyanate, in Sephadex gel beads permeated by aqueous solutions of these molecules. The molecular weights of these fractions were between 5400 and 200 000 and measurements of their diffusion coefficients inside a gel bead (
D) and in the free solution (
D
0), were performed using the fluorescence recovery after photobleaching method (FRAP). We also determined the coefficient of partitioning (
K
av) of these fractions between the gel and the free solvent, with a new microfluorimetric method. We found that, for Sephadex G-50, G-75, G-100, G-150 and G-200 gels,
K
av varied with the Stokes radius (
r
s) of the dextran molecules, in agreement with the formula of Laurent and Killander (J. Chromatogr. 14 (1964) 317). For Sephadex G-100, G-150 and G-200 gels,
D/
D
0 varied with r
s according to the theory of Ogston et al. (Proc. R. Soc. Lond. 333 (1973) 297). In addition, these theories predict a relation linking
D/
D
0 to
K
av which was wen verified. Our work is the first systematic study of the translational diffusion of macromolecules in a chromatography gel. These measurements should allow a better evaluation of the factors which influence the resolution in exclusion chromatography. In addition, the diffusion of macromolecules in gels may provide models for the diffusion of these molecules in the cytoplasm of living cells and in connective biological tissues.</description><identifier>ISSN: 0301-4622</identifier><identifier>EISSN: 1873-4200</identifier><identifier>DOI: 10.1016/0301-4622(88)80030-9</identifier><identifier>PMID: 2466497</identifier><identifier>CODEN: BICIAZ</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>Biological and medical sciences ; Biopolymers ; Chromatography, Gel - methods ; Conformational dynamics in molecular biology ; Dextrans ; Diffusion ; Fluorescein isothiocyanate dextran ; Fluorescein-5-isothiocyanate - analogs & derivatives ; Fluoresceins ; Fundamental and applied biological sciences. Psychology ; Gels ; macromolecules ; Models, Theoretical ; Molecular biophysics ; Molecular Conformation ; Partition coefficient ; Sephadex gel ; Size-exclusion chromatography ; Spectrometry, Fluorescence ; Translational diffusion</subject><ispartof>Biophysical chemistry, 1988-09, Vol.31 (3), p.247-258</ispartof><rights>1988</rights><rights>1990 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-395e7b61cbcb3f070611a139036bd37cf7416c37d7484c8840dc8652114ab9593</citedby><cites>FETCH-LOGICAL-c417t-395e7b61cbcb3f070611a139036bd37cf7416c37d7484c8840dc8652114ab9593</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0301-4622(88)80030-9$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6771092$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2466497$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Poitevin, Eric</creatorcontrib><creatorcontrib>Wahl, Philippe</creatorcontrib><title>Study of the translational diffusion of macromolecules in beads of gel chromatography by the FRAP method</title><title>Biophysical chemistry</title><addtitle>Biophys Chem</addtitle><description>We measured the translational diffusion of fractions of dextrans labelled with fluorescein isothiocyanate, in Sephadex gel beads permeated by aqueous solutions of these molecules. The molecular weights of these fractions were between 5400 and 200 000 and measurements of their diffusion coefficients inside a gel bead (
D) and in the free solution (
D
0), were performed using the fluorescence recovery after photobleaching method (FRAP). We also determined the coefficient of partitioning (
K
av) of these fractions between the gel and the free solvent, with a new microfluorimetric method. We found that, for Sephadex G-50, G-75, G-100, G-150 and G-200 gels,
K
av varied with the Stokes radius (
r
s) of the dextran molecules, in agreement with the formula of Laurent and Killander (J. Chromatogr. 14 (1964) 317). For Sephadex G-100, G-150 and G-200 gels,
D/
D
0 varied with r
s according to the theory of Ogston et al. (Proc. R. Soc. Lond. 333 (1973) 297). In addition, these theories predict a relation linking
D/
D
0 to
K
av which was wen verified. Our work is the first systematic study of the translational diffusion of macromolecules in a chromatography gel. These measurements should allow a better evaluation of the factors which influence the resolution in exclusion chromatography. In addition, the diffusion of macromolecules in gels may provide models for the diffusion of these molecules in the cytoplasm of living cells and in connective biological tissues.</description><subject>Biological and medical sciences</subject><subject>Biopolymers</subject><subject>Chromatography, Gel - methods</subject><subject>Conformational dynamics in molecular biology</subject><subject>Dextrans</subject><subject>Diffusion</subject><subject>Fluorescein isothiocyanate dextran</subject><subject>Fluorescein-5-isothiocyanate - analogs & derivatives</subject><subject>Fluoresceins</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gels</subject><subject>macromolecules</subject><subject>Models, Theoretical</subject><subject>Molecular biophysics</subject><subject>Molecular Conformation</subject><subject>Partition coefficient</subject><subject>Sephadex gel</subject><subject>Size-exclusion chromatography</subject><subject>Spectrometry, Fluorescence</subject><subject>Translational diffusion</subject><issn>0301-4622</issn><issn>1873-4200</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkd-LFSEUxyWK7bb1HxT4EFEPUzo66rwEy9JWsFD041kcPbNjOONNneD-9zl7L_exfNHD9-Ph8DkIPafkLSVUvCOM0IaLtn2t1BtFatn0D9COKska3hLyEO3OyGP0JOdfpJ4KXqCLlgvBe7lD0_eyugOOIy4T4JLMkoMpPi4mYOfHcc31vcWzsSnOMYBdA2TsFzyAcXmL7iBgO9XUlHiXzH464OFw3-_m29VXPEOZonuKHo0mZHh2ui_Rz5sPP64_NbdfPn6-vrptLKeyNKzvQA6C2sEObCSSCEoNZT1hYnBM2lFyKiyTTnLFrVKcOKtE11LKzdB3PbtEr4599yn-XiEXPftsIQSzQFyzlkqSriXyvyDtKGtJryrIj2AVkHOCUe-Tn006aEr0tgm9adabZq2Uvt-E3gZ5ceq_DjO486eT-pq_POUmWxPGqt76fMaElJT0bcXeHzGo0v54SDpbD4sF5xPYol30_57jL8Jzo3Q</recordid><startdate>19880901</startdate><enddate>19880901</enddate><creator>Poitevin, Eric</creator><creator>Wahl, Philippe</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M81</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19880901</creationdate><title>Study of the translational diffusion of macromolecules in beads of gel chromatography by the FRAP method</title><author>Poitevin, Eric ; Wahl, Philippe</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-395e7b61cbcb3f070611a139036bd37cf7416c37d7484c8840dc8652114ab9593</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>Biological and medical sciences</topic><topic>Biopolymers</topic><topic>Chromatography, Gel - methods</topic><topic>Conformational dynamics in molecular biology</topic><topic>Dextrans</topic><topic>Diffusion</topic><topic>Fluorescein isothiocyanate dextran</topic><topic>Fluorescein-5-isothiocyanate - analogs & derivatives</topic><topic>Fluoresceins</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gels</topic><topic>macromolecules</topic><topic>Models, Theoretical</topic><topic>Molecular biophysics</topic><topic>Molecular Conformation</topic><topic>Partition coefficient</topic><topic>Sephadex gel</topic><topic>Size-exclusion chromatography</topic><topic>Spectrometry, Fluorescence</topic><topic>Translational diffusion</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Poitevin, Eric</creatorcontrib><creatorcontrib>Wahl, Philippe</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 3</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biophysical chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Poitevin, Eric</au><au>Wahl, Philippe</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Study of the translational diffusion of macromolecules in beads of gel chromatography by the FRAP method</atitle><jtitle>Biophysical chemistry</jtitle><addtitle>Biophys Chem</addtitle><date>1988-09-01</date><risdate>1988</risdate><volume>31</volume><issue>3</issue><spage>247</spage><epage>258</epage><pages>247-258</pages><issn>0301-4622</issn><eissn>1873-4200</eissn><coden>BICIAZ</coden><abstract>We measured the translational diffusion of fractions of dextrans labelled with fluorescein isothiocyanate, in Sephadex gel beads permeated by aqueous solutions of these molecules. The molecular weights of these fractions were between 5400 and 200 000 and measurements of their diffusion coefficients inside a gel bead (
D) and in the free solution (
D
0), were performed using the fluorescence recovery after photobleaching method (FRAP). We also determined the coefficient of partitioning (
K
av) of these fractions between the gel and the free solvent, with a new microfluorimetric method. We found that, for Sephadex G-50, G-75, G-100, G-150 and G-200 gels,
K
av varied with the Stokes radius (
r
s) of the dextran molecules, in agreement with the formula of Laurent and Killander (J. Chromatogr. 14 (1964) 317). For Sephadex G-100, G-150 and G-200 gels,
D/
D
0 varied with r
s according to the theory of Ogston et al. (Proc. R. Soc. Lond. 333 (1973) 297). In addition, these theories predict a relation linking
D/
D
0 to
K
av which was wen verified. Our work is the first systematic study of the translational diffusion of macromolecules in a chromatography gel. These measurements should allow a better evaluation of the factors which influence the resolution in exclusion chromatography. In addition, the diffusion of macromolecules in gels may provide models for the diffusion of these molecules in the cytoplasm of living cells and in connective biological tissues.</abstract><cop>London</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>2466497</pmid><doi>10.1016/0301-4622(88)80030-9</doi><tpages>12</tpages></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0301-4622 |
ispartof | Biophysical chemistry, 1988-09, Vol.31 (3), p.247-258 |
issn | 0301-4622 1873-4200 |
language | eng |
recordid | cdi_proquest_miscellaneous_78705207 |
source | MEDLINE; Elsevier ScienceDirect Journals Complete |
subjects | Biological and medical sciences Biopolymers Chromatography, Gel - methods Conformational dynamics in molecular biology Dextrans Diffusion Fluorescein isothiocyanate dextran Fluorescein-5-isothiocyanate - analogs & derivatives Fluoresceins Fundamental and applied biological sciences. Psychology Gels macromolecules Models, Theoretical Molecular biophysics Molecular Conformation Partition coefficient Sephadex gel Size-exclusion chromatography Spectrometry, Fluorescence Translational diffusion |
title | Study of the translational diffusion of macromolecules in beads of gel chromatography by the FRAP method |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-24T13%3A41%3A20IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Study%20of%20the%20translational%20diffusion%20of%20macromolecules%20in%20beads%20of%20gel%20chromatography%20by%20the%20FRAP%20method&rft.jtitle=Biophysical%20chemistry&rft.au=Poitevin,%20Eric&rft.date=1988-09-01&rft.volume=31&rft.issue=3&rft.spage=247&rft.epage=258&rft.pages=247-258&rft.issn=0301-4622&rft.eissn=1873-4200&rft.coden=BICIAZ&rft_id=info:doi/10.1016/0301-4622(88)80030-9&rft_dat=%3Cproquest_cross%3E15132098%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=15132098&rft_id=info:pmid/2466497&rft_els_id=0301462288800309&rfr_iscdi=true |