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000153465 037__ $$aDZNE-2020-01462
000153465 041__ $$aEnglish
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000153465 1001_ $$00000-0003-3770-2594$$aKoo, Chek Ziu$$b0
000153465 245__ $$aThe tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex
000153465 260__ $$aBethesda, Md.$$bSoc.$$c2020
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000153465 520__ $$aA disintegrin and metalloprotease 10 (ADAM10) is a transmembrane protein essential for embryonic development, and its dysregulation underlies disorders such as cancer, Alzheimer's disease, and inflammation. ADAM10 is a "molecular scissor" that proteolytically cleaves the extracellular region from >100 substrates, including Notch, amyloid precursor protein, cadherins, growth factors, and chemokines. ADAM10 has been recently proposed to function as six distinct scissors with different substrates, depending on its association with one of six regulatory tetraspanins, termed TspanC8s. However, it remains unclear to what degree ADAM10 function critically depends on a TspanC8 partner, and a lack of monoclonal antibodies specific for most TspanC8s has hindered investigation of this question. To address this knowledge gap, here we designed an immunogen to generate the first monoclonal antibodies targeting Tspan15, a model TspanC8. The immunogen was created in an ADAM10-knockout mouse cell line stably overexpressing human Tspan15, because we hypothesized that expression in this cell line would expose epitopes that are normally blocked by ADAM10. Following immunization of mice, this immunogen strategy generated four Tspan15 antibodies. Using these antibodies, we show that endogenous Tspan15 and ADAM10 co-localize on the cell surface, that ADAM10 is the principal Tspan15-interacting protein, that endogenous Tspan15 expression requires ADAM10 in cell lines and primary cells, and that a synthetic ADAM10/Tspan15 fusion protein is a functional scissor. Furthermore, two of the four antibodies impaired ADAM10/Tspan15 activity. These findings suggest that Tspan15 directly interacts with ADAM10 in a functional scissor complex.
000153465 536__ $$0G:(DE-HGF)POF3-342$$a342 - Disease Mechanisms and Model Systems (POF3-342)$$cPOF3-342$$fPOF III$$x0
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000153465 650_2 $$2MeSH$$aA549 Cells
000153465 650_2 $$2MeSH$$aADAM10 Protein: genetics
000153465 650_2 $$2MeSH$$aADAM10 Protein: metabolism
000153465 650_2 $$2MeSH$$aAmyloid Precursor Protein Secretases: genetics
000153465 650_2 $$2MeSH$$aAmyloid Precursor Protein Secretases: metabolism
000153465 650_2 $$2MeSH$$aAnimals
000153465 650_2 $$2MeSH$$aHEK293 Cells
000153465 650_2 $$2MeSH$$aHumans
000153465 650_2 $$2MeSH$$aJurkat Cells
000153465 650_2 $$2MeSH$$aMembrane Proteins: genetics
000153465 650_2 $$2MeSH$$aMembrane Proteins: metabolism
000153465 650_2 $$2MeSH$$aMice
000153465 650_2 $$2MeSH$$aMice, Knockout
000153465 650_2 $$2MeSH$$aMultiprotein Complexes: genetics
000153465 650_2 $$2MeSH$$aMultiprotein Complexes: metabolism
000153465 650_2 $$2MeSH$$aTetraspanins: genetics
000153465 650_2 $$2MeSH$$aTetraspanins: metabolism
000153465 7001_ $$00000-0001-6821-4089$$aHarrison, Neale$$b1
000153465 7001_ $$aNoy, Peter J.$$b2
000153465 7001_ $$aSzyroka, Justyna$$b3
000153465 7001_ $$aMatthews, Alexandra L.$$b4
000153465 7001_ $$0P:(DE-2719)2811769$$aHsia, Hung-En$$b5$$udzne
000153465 7001_ $$0P:(DE-2719)2810938$$aMüller, Stephan A$$b6$$udzne
000153465 7001_ $$0P:(DE-2719)2812852$$aTüshaus, Johanna$$b7$$udzne
000153465 7001_ $$00000-0002-6227-4483$$aGoulding, Joelle$$b8
000153465 7001_ $$aWillis, Katie$$b9
000153465 7001_ $$aApicella, Clara$$b10
000153465 7001_ $$aCragoe, Bethany$$b11
000153465 7001_ $$aDavis, Edward$$b12
000153465 7001_ $$aKeles, Murat$$b13
000153465 7001_ $$aMalinova, Antonia$$b14
000153465 7001_ $$aMcFarlane, Thomas A.$$b15
000153465 7001_ $$aMorrison, Philip R.$$b16
000153465 7001_ $$aNguyen, Hanh T. H.$$b17
000153465 7001_ $$aSykes, Michael C.$$b18
000153465 7001_ $$aAhmed, Haroon$$b19
000153465 7001_ $$aDi Maio, Alessandro$$b20
000153465 7001_ $$aSeipold, Lisa$$b21
000153465 7001_ $$aSaftig, Paul$$b22
000153465 7001_ $$00000-0002-4309-4858$$aCull, Eleanor$$b23
000153465 7001_ $$aPliotas, Christos$$b24
000153465 7001_ $$aRubinstein, Eric$$b25
000153465 7001_ $$00000-0002-3187-2130$$aPoulter, Natalie S.$$b26
000153465 7001_ $$aBriddon, Stephen J.$$b27
000153465 7001_ $$aHolliday, Nicholas D.$$b28
000153465 7001_ $$0P:(DE-2719)2181459$$aLichtenthaler, Stefan$$b29$$udzne
000153465 7001_ $$00000-0002-1189-0091$$aTomlinson, Michael G.$$b30$$eCorresponding author
000153465 773__ $$0PERI:(DE-600)1474604-9$$a10.1074/jbc.RA120.012601$$gVol. 295, no. 36, p. 12822 - 12839$$n36$$p12822 - 12839$$tThe journal of biological chemistry$$v295$$x0021-9258$$y2020
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