Alternative Splicing of the Angiogenesis Associated Extra-Domain B of Fibronectin Regulates the Accessibility of the B-C Loop of the Type III Repeat 8 英文参考文献.docVIP

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Alternative Splicing of the Angiogenesis Associated Extra-Domain B of Fibronectin Regulates the Accessibility of the B-C Loop of the Type III Repeat 8 英文参考文献.doc

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Alternative Splicing of the Angiogenesis Associated Extra-Domain B of Fibronectin Regulates the Accessibility of the B-C Loop of the Type III Repeat 8 英文参考文献

AlternativeSplicingoftheAngiogenesisAssociated Extra-DomainBofFibronectinRegulatesthe AccessibilityoftheB-CLoopoftheTypeIIIRepeat8 ElisaVentura1,FrancescaSassi1,AriannaParodi1,EnricaBalza2,LauraBorsi2,PatriziaCastellani2, BarbaraCarnemolla3,LucianoZardi1* 1LaboratoryofRecombinantTherapeuticProteins,CBA–AdvancedBiotechnologyCentre,Genoa,Italy,2LaboratoryofCellBiology,IstitutoNazionaleperlaRicercasul Cancro,Genoa,Italy,3LaboratoryofImmunology,IstitutoNazionaleperlaRicercasulCancro,Genoa,Italy Abstract Background: Fibronectin (FN) is a multi-domain molecule involved in many cellular processes, including tissue repair, embryogenesis,bloodclotting,andcellmigration/adhesion.ThebiologicalactivitiesofFNaremediatedbyexposedloops locatedmainlyattheinterdomaininterfacesthatinteractwithvariousmoleculessuchas,butnotonly,integrins.Different FNisoformsarisefromthealternativesplicingofthepre-mRNA.Inmalignancies,thesplicingpatternofFNpre-mRNAis altered;inparticular,theFNisoformcontainingtheextra-domainB(ED-B),acompleteFNtypeIIIrepeatconstitutedby91 residues,isundetectableinnormaladulttissues,butexhibitsamuchgreaterexpressioninfetalandtumortissues,andis accumulatedaroundneovasculatureduringangiogenicprocesses,thusmakingED-Boneofthebestmarkersandtargetsof angiogenesis.ThefunctionsofED-Barestillunclear;however,ithasbeenpostulatedthattheinsertionofanextra-domain suchasED-Bmodifiesthedomain-domaininterfaceandmayunmaskloopsthatareotherwisecryptic,thusgivingFNnew potentialactivities. Methodology: We used the mAb C6, which reacts with ED-B containing FN, but not with ED-B-free FN and various recombinantFNfragmentscontainingmutations,topreciselylocalizetheepitopesrecognizedbythemAbC6. Conclusion:Weformallydemonstratedthattheinclusionofthealternativelysplicedangiogenesis-associatedED-Bleadsto the unmasking of the FNIII 8 B-C loop that is cryptic in FN molecules lacking ED-B. Thus, the mAb C6, in addition to providinganewreagentforangiogenesistargeting,representsanewtoolforthestudyofthepoten