線性泛素鏈組裝復(fù)合體LUBAC與整合素連接激酶ILK的相互作用鑒定與功能研究

線性泛素鏈組裝復(fù)合體LUBAC與整合素連接激酶ILK的相互作用鑒定與功能研究摘要：線性泛素鏈組裝復(fù)合體（LUBAC）是一種重要的信號(hào)轉(zhuǎn)導(dǎo)復(fù)合體，能夠通過形成鏈性泛素鏈來調(diào)節(jié)多個(gè)信號(hào)通路。在本研究中，我們發(fā)現(xiàn)LUBAC能夠與整合素連接激酶（ILK）發(fā)生特異性相互作用。通過表達(dá)和純化蛋白質(zhì)，我們鑒定到LUBAC中的HOIP子單位能夠結(jié)合于ILK的N末端結(jié)構(gòu)域，這種相互作用能夠調(diào)節(jié)ILK的酶活性和下游信號(hào)通路。實(shí)驗(yàn)結(jié)果表明，LUBAC/ILK相互作用可以影響膠原蛋白合成和細(xì)胞外基質(zhì)的合成。我們的研究揭示了LUBAC和ILK之間的新的信號(hào)通路，對(duì)于進(jìn)一步了解細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)機(jī)制具有重要意義。

關(guān)鍵詞：線性泛素鏈組裝復(fù)合體，整合素連接激酶，相互作用，酶活性，信號(hào)通路

Introduction

線性泛素鏈組裝復(fù)合體（LUBAC）是一種由三個(gè)亞單位HOIL-1L、HOIP和Sharpin組成的信號(hào)轉(zhuǎn)導(dǎo)復(fù)合體。它通過形成鏈性泛素鏈來調(diào)節(jié)多個(gè)信號(hào)通路，包括NF-kB通路、IRF3通路、細(xì)胞凋亡和血管生成等。近年來的一些研究表明，LUBAC還能夠參與到其他生物學(xué)過程中，如纖維化和癌癥發(fā)生等。而整合素連接激酶（ILK）是一種細(xì)胞外基質(zhì)信號(hào)傳導(dǎo)的重要調(diào)節(jié)因子，能夠通過參與細(xì)胞黏附、遷移、增殖等過程而產(chǎn)生生物學(xué)效應(yīng)。ILK的活性和下游信號(hào)通路的調(diào)節(jié)對(duì)于許多生理和病理情況都具有重要意義。因此，研究LUBAC與ILK之間的聯(lián)系對(duì)于深入了解細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)機(jī)制具有重要意義。

Results

我們通過大規(guī)模篩選鑒定到LUBAC能夠與ILK發(fā)生特異性相互作用，而這種相互作用主要是由LUBAC中的HOIP子單位和ILK的N末端結(jié)構(gòu)域之間的結(jié)合引起的。我們進(jìn)一步通過蛋白質(zhì)相互作用實(shí)驗(yàn)和熒光共聚焦顯微鏡來驗(yàn)證LUBAC和ILK之間的相互作用。我們發(fā)現(xiàn)，LUBAC能夠顯著地抑制ILK的活性，并通過抑制下游信號(hào)通路的激活來對(duì)細(xì)胞生物學(xué)活性產(chǎn)生影響。此外，我們還發(fā)現(xiàn)，LUBAC/ILK相互作用可以影響膠原蛋白的合成和細(xì)胞外基質(zhì)的生成，從而影響細(xì)胞黏附和遷移。

Conclusion

我們的研究揭示了LUBAC與ILK之間的新的信號(hào)通路，并證明了LUBAC與ILK之間的相互作用可以影響細(xì)胞生物學(xué)活性。因此，LUBAC和ILK的相互作用在細(xì)胞黏附、遷移、增殖等生物學(xué)過程中具有重要作用。這種相互作用為深入了解細(xì)胞信號(hào)調(diào)節(jié)機(jī)制提供了新思路，同時(shí)也可能在未來的治療策略中發(fā)揮一定的作用Furthermore,ourstudyhighlightstheimportanceoftheHOIPsubunitofLUBACinmediatingtheinteractionwithILKanditsdownstreameffectsoncellbiology.TheinhibitionofILKactivitybyLUBACsuggestsapotentialtherapeuticstrategyfordiseasesassociatedwithdysregulatedILKsignaling,suchascancerandcardiovasculardiseases.

Inconclusion,ourfindingsprovidenovelinsightsintothesignalingpathwaysinvolvingLUBACandILKandtheirimpactoncellularbehavior.TheunderstandingoftheinterplaybetweenLUBACandILKcancontributetothedevelopmentofnewapproachestotreatvariouspathologicalconditions.FutureresearchisnecessarytoexploretheprecisedetailsoftheLUBAC/ILKsignalingaxisanditspotentialasatherapeutictargetAdditionally,furtherinvestigationisnecessarytodeterminethefunctionalsignificanceoftheLUBAC/ILKsignalingaxisinvariouspathologicalconditions.Theidentificationofdownstreameffectorsofthispathwaycanprovidepotentialtargetsfordrugdevelopmentandtherapeuticinterventions.

Moreover,theroleofLUBACinregulatingimmuneresponsesandinflammationremainstobefullyelucidated.ItisknownthatLUBACisinvolvedintheactivationofNF-κBsignaling,amajortranscriptionalregulatoroftheimmunesystem.FuturestudiesmayrevealhowLUBAC/ILKsignalingcontributestoinflammationandimmuneresponsesandwhethertargetingthispathwaycanhavetherapeuticimplicationsforautoimmuneandinflammatorydiseases.

Furthermore,thecrosstalkbetweenLUBAC/ILKsignalingandothersignalingpathways,suchasthePI3K/AKTandMAPKpathways,needstobeexplored.Thesepathwaysareknowntobederegulatedinvariousdiseases,includingcancerandcardiovasculardiseases,andoftenintersectwitheachother.ElucidatingtheinteractionsbetweenLUBAC/ILKsignalingandthesepathwayscanprovideamorecomprehensiveunderstandingoftheunderlyingmechanismsofdiseasepathogenesis.

Insummary,theLUBAC/ILKsignalingaxisrepresentsacomplexandintriguingpathwaywithdiverserolesincellularbehavioranddiseasepathogenesis.AbetterunderstandingofthispathwaycanpavethewayforthedevelopmentofnoveltherapeuticstrategiesforvariouspathologicalconditionsTheLUBAC/ILKsignalingaxishasalsobeenimplicatedinvariousotherpathologicalconditionsbeyondcancerandimmunedisorders.Onesuchconditioniscardiovasculardisease.StudieshaveshownthatLUBACisinvolvedinregulatingendothelialcellfunction,whichplaysacriticalroleinmaintainingcardiovascularhomeostasis.LUBAC-mediatedactivationofNF-κBhasbeenshowntopromotetheexpressionofpro-inflammatorycytokinesandadhesionmoleculesinendothelialcells,therebycontributingtothedevelopmentofatherosclerosis.ILKhasalsobeenshowntoplayaroleinvascularsmoothmusclecellproliferationandmigration,whicharekeyeventsinthepathogenesisofrestenosisfollowingangioplasty.TargetingtheLUBAC/ILKsignalingaxismaythereforerepresentapotentialtherapeuticstrategyforcardiovasculardisease.

AnotherconditioninwhichtheLUBAC/ILKsignalingaxishasbeenimplicatedisneurologicaldisorders.StudieshaveshownthatLUBACisinvolvedinregulatingneuronalsurvivalandsynapticplasticity.LUBAC-mediatedactivationofNF-κBhasbeenshowntopromotetheexpressionofpro-inflammatorycytokinesandoxidativestressinneurons,whichcontributestoneuronallossanddysfunctioninconditionssuchasAlzheimer'sdiseaseandParkinson'sdisease.ILKhasalsobeenshowntoplayaroleinpromotingdendriticspineformationandsynapticplasticity,whicharecriticaleventsinlearningandmemory.TargetingtheLUBAC/ILKsignalingaxismaythereforerepresentapotentialtherapeuticstrategyforneurologicaldisorders.

Inconclusion,theLUBAC/ILKsignalingaxisrepresentsacomplexandmultifunctionalpathwaythatplaysdiverserolesincellularbehavioranddiseasepathogenesis.WhilemuchprogresshasbeenmadeinunderstandingthemechanismsunderlyingLUBAC/ILKsignaling,manyquestionsstillremainunanswered,particularlyintermsoftheinteractionsbetweenthispathwayandothersignalingpathways.Abetterunderstandingofthispathwayiscrucialforthedevelopmentofnoveltherapeuticstrategiesforvariouspathologicalconditions,includingcancer,immunedisorders,cardiovasculardisease,an