A Six-Amino Acid Deletion in Basic Fibroblast Growth Factor Dissociates its Mitogenic Activity from its Plasminogen Activator-Inducing Capacity

A recombinant deletion mutant of the 155-amino acid form of human basic fibroblast growth factor (bFGF), lacking amino acid residues 27-32 (Lys-Asp-Pro-Lys-Arg-Leu), was expressed in Escherichia coli and purified to homogeneity by heparin-Sepharose affinity chromatography. When maintained in the pre...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS 1991-04, Vol.88 (7), p.2628-2632
Hauptverfasser: Isacchi, Antonella, Statuto, Massimo, Chiesa, Roberta, Bergonzoni, Laura, Rusnati, Marco, Sarmientos, Paolo, Ragnotti, Giovanni, Presta, Marco
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 2632
container_issue 7
container_start_page 2628
container_title Proceedings of the National Academy of Sciences - PNAS
container_volume 88
creator Isacchi, Antonella
Statuto, Massimo
Chiesa, Roberta
Bergonzoni, Laura
Rusnati, Marco
Sarmientos, Paolo
Ragnotti, Giovanni
Presta, Marco
description A recombinant deletion mutant of the 155-amino acid form of human basic fibroblast growth factor (bFGF), lacking amino acid residues 27-32 (Lys-Asp-Pro-Lys-Arg-Leu), was expressed in Escherichia coli and purified to homogeneity by heparin-Sepharose affinity chromatography. When maintained in the presence of an equimolar concentration of soluble heparin, the bFGF mutant (M1-bFGF) is as potent as bFGF in stimulating cell proliferation in normal and transformed fetal bovine aortic endothelial cells, in adult bovine aortic endothelial cells, and in NIH 3T3 fibroblasts. However, under the same experimental conditions, M1-bFGF is at least 100 times less efficient than bFGF in stimulating plasminogen activator (PA) production in endothelial cells, as assayed by chromogenic PA assay, SDS/PAGE zymography, and Northern blot analysis of urokinase-type PA mRNA. In the presence of heparin, M1-bFGF binds to bFGF plasma membrane receptors present on endothelial cells in a manner undistinguishable from bFGF. It also induces the same tyrosine phosphorylation pattern when added to NIH 3T3 cells. The data suggest that the PA-inducing activity of bFGF may depend upon a functional domain that differs from those involved in the mitogenic activity of the growth factor and that the binding of bFGF to its plasma membrane receptor may not be sufficient to induce urokinase-type PA production in endothelial cells.
doi_str_mv 10.1073/pnas.88.7.2628
format Article
fullrecord <record><control><sourceid>jstor_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_51291</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><jstor_id>2356238</jstor_id><sourcerecordid>2356238</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3728-97d10df2c79a0667585fad598dec0097c2f9060d308489e90aee51a6f41147d83</originalsourceid><addsrcrecordid>eNptkU9PHCEYh4mxsVvt1VNNOPU2U2D-AImX6dq1JhpN2p4JC8yKmYUNsFY_Rb9ymY5dNfHE4Xmf3xveHwDHGJUY0erLxslYMlbSkrSE7YEZRhwXbc3RPpghRGjBalK_Bx9ivEMI8YahA3CAWc1Jy2fgTwd_2IeiW1vnYaeshmdmMMl6B62DX2W0Ci7sMvjlIGOC58H_TrdwIVXyAZ7ZGL2yMpkIbYrwyia_Mi4rnUr23qZH2Ae__sdusj8uyXyiMicUF05vlXUrOJcbqbJwBN71cojm49N7CH4tvv2cfy8ur88v5t1loSpKWMGpxkj3RFEuUdvShjW91A1n2qj8SapIz1GLdIVYzbjhSBrTYNn2NcY11aw6BKdT7ma7XButjEtBDmIT7FqGR-GlFa-Js7di5e9FgwnHWS8nXQUfYzD9zsRIjL2IsRfBmKBi7CULJy_3PY9PRWT--YmP3n-680W_HYZkHtKLoDcHM_808buYD7wbIFXTkopVfwFBzq2c</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype></control><display><type>article</type><title>A Six-Amino Acid Deletion in Basic Fibroblast Growth Factor Dissociates its Mitogenic Activity from its Plasminogen Activator-Inducing Capacity</title><source>MEDLINE</source><source>JSTOR Archive Collection A-Z Listing</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><source>Free Full-Text Journals in Chemistry</source><creator>Isacchi, Antonella ; Statuto, Massimo ; Chiesa, Roberta ; Bergonzoni, Laura ; Rusnati, Marco ; Sarmientos, Paolo ; Ragnotti, Giovanni ; Presta, Marco</creator><creatorcontrib>Isacchi, Antonella ; Statuto, Massimo ; Chiesa, Roberta ; Bergonzoni, Laura ; Rusnati, Marco ; Sarmientos, Paolo ; Ragnotti, Giovanni ; Presta, Marco</creatorcontrib><description>A recombinant deletion mutant of the 155-amino acid form of human basic fibroblast growth factor (bFGF), lacking amino acid residues 27-32 (Lys-Asp-Pro-Lys-Arg-Leu), was expressed in Escherichia coli and purified to homogeneity by heparin-Sepharose affinity chromatography. When maintained in the presence of an equimolar concentration of soluble heparin, the bFGF mutant (M1-bFGF) is as potent as bFGF in stimulating cell proliferation in normal and transformed fetal bovine aortic endothelial cells, in adult bovine aortic endothelial cells, and in NIH 3T3 fibroblasts. However, under the same experimental conditions, M1-bFGF is at least 100 times less efficient than bFGF in stimulating plasminogen activator (PA) production in endothelial cells, as assayed by chromogenic PA assay, SDS/PAGE zymography, and Northern blot analysis of urokinase-type PA mRNA. In the presence of heparin, M1-bFGF binds to bFGF plasma membrane receptors present on endothelial cells in a manner undistinguishable from bFGF. It also induces the same tyrosine phosphorylation pattern when added to NIH 3T3 cells. The data suggest that the PA-inducing activity of bFGF may depend upon a functional domain that differs from those involved in the mitogenic activity of the growth factor and that the binding of bFGF to its plasma membrane receptor may not be sufficient to induce urokinase-type PA production in endothelial cells.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.88.7.2628</identifier><identifier>PMID: 1849269</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>3T3 cells ; Amino Acid Sequence ; Amino acids ; Base Sequence ; Binding sites ; Blotting, Northern ; Cell Division - drug effects ; Cell growth ; Cell Line ; Cell membranes ; Chromosome Deletion ; DNA Replication - drug effects ; Down-Regulation ; Endothelial cells ; Fibroblast Growth Factor 2 - genetics ; Fibroblast Growth Factor 2 - isolation &amp; purification ; Fibroblast Growth Factor 2 - metabolism ; Fibroblast Growth Factor 2 - pharmacology ; Genes, Synthetic ; Heparin ; Humans ; Kinetics ; Mitogens ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; NIH 3T3 cells ; Oligonucleotide Probes ; Phosphorylation ; Plasmids ; Plasminogen Activators ; Receptors ; Receptors, Cell Surface - drug effects ; Receptors, Cell Surface - metabolism ; Receptors, Fibroblast Growth Factor</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1991-04, Vol.88 (7), p.2628-2632</ispartof><rights>Copyright 1991 The National Academy of Sciences of the United States of America</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3728-97d10df2c79a0667585fad598dec0097c2f9060d308489e90aee51a6f41147d83</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/88/7.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/2356238$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/2356238$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,727,780,784,803,885,27924,27925,53791,53793,58017,58250</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1849269$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Isacchi, Antonella</creatorcontrib><creatorcontrib>Statuto, Massimo</creatorcontrib><creatorcontrib>Chiesa, Roberta</creatorcontrib><creatorcontrib>Bergonzoni, Laura</creatorcontrib><creatorcontrib>Rusnati, Marco</creatorcontrib><creatorcontrib>Sarmientos, Paolo</creatorcontrib><creatorcontrib>Ragnotti, Giovanni</creatorcontrib><creatorcontrib>Presta, Marco</creatorcontrib><title>A Six-Amino Acid Deletion in Basic Fibroblast Growth Factor Dissociates its Mitogenic Activity from its Plasminogen Activator-Inducing Capacity</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>A recombinant deletion mutant of the 155-amino acid form of human basic fibroblast growth factor (bFGF), lacking amino acid residues 27-32 (Lys-Asp-Pro-Lys-Arg-Leu), was expressed in Escherichia coli and purified to homogeneity by heparin-Sepharose affinity chromatography. When maintained in the presence of an equimolar concentration of soluble heparin, the bFGF mutant (M1-bFGF) is as potent as bFGF in stimulating cell proliferation in normal and transformed fetal bovine aortic endothelial cells, in adult bovine aortic endothelial cells, and in NIH 3T3 fibroblasts. However, under the same experimental conditions, M1-bFGF is at least 100 times less efficient than bFGF in stimulating plasminogen activator (PA) production in endothelial cells, as assayed by chromogenic PA assay, SDS/PAGE zymography, and Northern blot analysis of urokinase-type PA mRNA. In the presence of heparin, M1-bFGF binds to bFGF plasma membrane receptors present on endothelial cells in a manner undistinguishable from bFGF. It also induces the same tyrosine phosphorylation pattern when added to NIH 3T3 cells. The data suggest that the PA-inducing activity of bFGF may depend upon a functional domain that differs from those involved in the mitogenic activity of the growth factor and that the binding of bFGF to its plasma membrane receptor may not be sufficient to induce urokinase-type PA production in endothelial cells.</description><subject>3T3 cells</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Base Sequence</subject><subject>Binding sites</subject><subject>Blotting, Northern</subject><subject>Cell Division - drug effects</subject><subject>Cell growth</subject><subject>Cell Line</subject><subject>Cell membranes</subject><subject>Chromosome Deletion</subject><subject>DNA Replication - drug effects</subject><subject>Down-Regulation</subject><subject>Endothelial cells</subject><subject>Fibroblast Growth Factor 2 - genetics</subject><subject>Fibroblast Growth Factor 2 - isolation &amp; purification</subject><subject>Fibroblast Growth Factor 2 - metabolism</subject><subject>Fibroblast Growth Factor 2 - pharmacology</subject><subject>Genes, Synthetic</subject><subject>Heparin</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Mitogens</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Site-Directed</subject><subject>NIH 3T3 cells</subject><subject>Oligonucleotide Probes</subject><subject>Phosphorylation</subject><subject>Plasmids</subject><subject>Plasminogen Activators</subject><subject>Receptors</subject><subject>Receptors, Cell Surface - drug effects</subject><subject>Receptors, Cell Surface - metabolism</subject><subject>Receptors, Fibroblast Growth Factor</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkU9PHCEYh4mxsVvt1VNNOPU2U2D-AImX6dq1JhpN2p4JC8yKmYUNsFY_Rb9ymY5dNfHE4Xmf3xveHwDHGJUY0erLxslYMlbSkrSE7YEZRhwXbc3RPpghRGjBalK_Bx9ivEMI8YahA3CAWc1Jy2fgTwd_2IeiW1vnYaeshmdmMMl6B62DX2W0Ci7sMvjlIGOC58H_TrdwIVXyAZ7ZGL2yMpkIbYrwyia_Mi4rnUr23qZH2Ae__sdusj8uyXyiMicUF05vlXUrOJcbqbJwBN71cojm49N7CH4tvv2cfy8ur88v5t1loSpKWMGpxkj3RFEuUdvShjW91A1n2qj8SapIz1GLdIVYzbjhSBrTYNn2NcY11aw6BKdT7ma7XButjEtBDmIT7FqGR-GlFa-Js7di5e9FgwnHWS8nXQUfYzD9zsRIjL2IsRfBmKBi7CULJy_3PY9PRWT--YmP3n-680W_HYZkHtKLoDcHM_808buYD7wbIFXTkopVfwFBzq2c</recordid><startdate>19910401</startdate><enddate>19910401</enddate><creator>Isacchi, Antonella</creator><creator>Statuto, Massimo</creator><creator>Chiesa, Roberta</creator><creator>Bergonzoni, Laura</creator><creator>Rusnati, Marco</creator><creator>Sarmientos, Paolo</creator><creator>Ragnotti, Giovanni</creator><creator>Presta, Marco</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><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>5PM</scope></search><sort><creationdate>19910401</creationdate><title>A Six-Amino Acid Deletion in Basic Fibroblast Growth Factor Dissociates its Mitogenic Activity from its Plasminogen Activator-Inducing Capacity</title><author>Isacchi, Antonella ; Statuto, Massimo ; Chiesa, Roberta ; Bergonzoni, Laura ; Rusnati, Marco ; Sarmientos, Paolo ; Ragnotti, Giovanni ; Presta, Marco</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3728-97d10df2c79a0667585fad598dec0097c2f9060d308489e90aee51a6f41147d83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>3T3 cells</topic><topic>Amino Acid Sequence</topic><topic>Amino acids</topic><topic>Base Sequence</topic><topic>Binding sites</topic><topic>Blotting, Northern</topic><topic>Cell Division - drug effects</topic><topic>Cell growth</topic><topic>Cell Line</topic><topic>Cell membranes</topic><topic>Chromosome Deletion</topic><topic>DNA Replication - drug effects</topic><topic>Down-Regulation</topic><topic>Endothelial cells</topic><topic>Fibroblast Growth Factor 2 - genetics</topic><topic>Fibroblast Growth Factor 2 - isolation &amp; purification</topic><topic>Fibroblast Growth Factor 2 - metabolism</topic><topic>Fibroblast Growth Factor 2 - pharmacology</topic><topic>Genes, Synthetic</topic><topic>Heparin</topic><topic>Humans</topic><topic>Kinetics</topic><topic>Mitogens</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Site-Directed</topic><topic>NIH 3T3 cells</topic><topic>Oligonucleotide Probes</topic><topic>Phosphorylation</topic><topic>Plasmids</topic><topic>Plasminogen Activators</topic><topic>Receptors</topic><topic>Receptors, Cell Surface - drug effects</topic><topic>Receptors, Cell Surface - metabolism</topic><topic>Receptors, Fibroblast Growth Factor</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Isacchi, Antonella</creatorcontrib><creatorcontrib>Statuto, Massimo</creatorcontrib><creatorcontrib>Chiesa, Roberta</creatorcontrib><creatorcontrib>Bergonzoni, Laura</creatorcontrib><creatorcontrib>Rusnati, Marco</creatorcontrib><creatorcontrib>Sarmientos, Paolo</creatorcontrib><creatorcontrib>Ragnotti, Giovanni</creatorcontrib><creatorcontrib>Presta, Marco</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Isacchi, Antonella</au><au>Statuto, Massimo</au><au>Chiesa, Roberta</au><au>Bergonzoni, Laura</au><au>Rusnati, Marco</au><au>Sarmientos, Paolo</au><au>Ragnotti, Giovanni</au><au>Presta, Marco</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Six-Amino Acid Deletion in Basic Fibroblast Growth Factor Dissociates its Mitogenic Activity from its Plasminogen Activator-Inducing Capacity</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1991-04-01</date><risdate>1991</risdate><volume>88</volume><issue>7</issue><spage>2628</spage><epage>2632</epage><pages>2628-2632</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>A recombinant deletion mutant of the 155-amino acid form of human basic fibroblast growth factor (bFGF), lacking amino acid residues 27-32 (Lys-Asp-Pro-Lys-Arg-Leu), was expressed in Escherichia coli and purified to homogeneity by heparin-Sepharose affinity chromatography. When maintained in the presence of an equimolar concentration of soluble heparin, the bFGF mutant (M1-bFGF) is as potent as bFGF in stimulating cell proliferation in normal and transformed fetal bovine aortic endothelial cells, in adult bovine aortic endothelial cells, and in NIH 3T3 fibroblasts. However, under the same experimental conditions, M1-bFGF is at least 100 times less efficient than bFGF in stimulating plasminogen activator (PA) production in endothelial cells, as assayed by chromogenic PA assay, SDS/PAGE zymography, and Northern blot analysis of urokinase-type PA mRNA. In the presence of heparin, M1-bFGF binds to bFGF plasma membrane receptors present on endothelial cells in a manner undistinguishable from bFGF. It also induces the same tyrosine phosphorylation pattern when added to NIH 3T3 cells. The data suggest that the PA-inducing activity of bFGF may depend upon a functional domain that differs from those involved in the mitogenic activity of the growth factor and that the binding of bFGF to its plasma membrane receptor may not be sufficient to induce urokinase-type PA production in endothelial cells.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>1849269</pmid><doi>10.1073/pnas.88.7.2628</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0027-8424
ispartof Proceedings of the National Academy of Sciences - PNAS, 1991-04, Vol.88 (7), p.2628-2632
issn 0027-8424
1091-6490
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_51291
source MEDLINE; JSTOR Archive Collection A-Z Listing; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry
subjects 3T3 cells
Amino Acid Sequence
Amino acids
Base Sequence
Binding sites
Blotting, Northern
Cell Division - drug effects
Cell growth
Cell Line
Cell membranes
Chromosome Deletion
DNA Replication - drug effects
Down-Regulation
Endothelial cells
Fibroblast Growth Factor 2 - genetics
Fibroblast Growth Factor 2 - isolation & purification
Fibroblast Growth Factor 2 - metabolism
Fibroblast Growth Factor 2 - pharmacology
Genes, Synthetic
Heparin
Humans
Kinetics
Mitogens
Molecular Sequence Data
Mutagenesis, Site-Directed
NIH 3T3 cells
Oligonucleotide Probes
Phosphorylation
Plasmids
Plasminogen Activators
Receptors
Receptors, Cell Surface - drug effects
Receptors, Cell Surface - metabolism
Receptors, Fibroblast Growth Factor
title A Six-Amino Acid Deletion in Basic Fibroblast Growth Factor Dissociates its Mitogenic Activity from its Plasminogen Activator-Inducing Capacity
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T15%3A25%3A57IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-jstor_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20Six-Amino%20Acid%20Deletion%20in%20Basic%20Fibroblast%20Growth%20Factor%20Dissociates%20its%20Mitogenic%20Activity%20from%20its%20Plasminogen%20Activator-Inducing%20Capacity&rft.jtitle=Proceedings%20of%20the%20National%20Academy%20of%20Sciences%20-%20PNAS&rft.au=Isacchi,%20Antonella&rft.date=1991-04-01&rft.volume=88&rft.issue=7&rft.spage=2628&rft.epage=2632&rft.pages=2628-2632&rft.issn=0027-8424&rft.eissn=1091-6490&rft_id=info:doi/10.1073/pnas.88.7.2628&rft_dat=%3Cjstor_pubme%3E2356238%3C/jstor_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_id=info:pmid/1849269&rft_jstor_id=2356238&rfr_iscdi=true