title: Control of ADAM17 activity by regulation of its cellular localisation.
abstract: An important, irreversible step in many signalling pathways is the shedding of membrane-anchored proteins. A Disintegrin And Metalloproteinase (ADAM) 17 is one of the major sheddases involved in a variety of physiological and pathophysiological processes including regeneration, differentiation, and cancer progression. This central role in signalling implies that ADAM17 activity has to be tightly regulated, including at the level of localisation. Most mature ADAM17 is localised intracellularly, with only a small amount at the cell surface. We found that ADAM17 is constitutively internalised by clathrin-coated pits and that physiological stimulators such as GPCR ligands induce ADAM17-mediated shedding, but do not alter the cell-surface abundance of the protease. In contrast, the PKC-activating phorbol ester PMA, often used as a strong inducer of ADAM17, causes not only proteolysis by ADAM17 but also a rapid increase of the mature protease at the cell surface. This is followed by internalisation and subsequent degradation of the protease. Eventually, this leads to a substantial downregulation of mature ADAM17. Our results therefore imply that physiological activation of ADAM17 does not rely on its relocalisation, but that PMA-induced PKC activity drastically dysregulates the localisation of ADAM17.
authors.affiliation: Institute of Biochemistry, Kiel University, Olshausenstr 40, 24098 Kiel, Germany.
authors.forename: Inken
authors.initials: I
authors.lastname: Lorenzen
authors.affiliation: Institute of Biochemistry, Kiel University, Olshausenstr 40, 24098 Kiel, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Lokau
authors.affiliation: Institute of Biochemistry, Kiel University, Olshausenstr 40, 24098 Kiel, Germany.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Korpys
authors.affiliation: Institute of Biochemistry, Kiel University, Olshausenstr 40, 24098 Kiel, Germany.
authors.forename: Mirja
authors.initials: M
authors.lastname: Oldefest
authors.affiliation: Institute of Biochemistry, Kiel University, Olshausenstr 40, 24098 Kiel, Germany.
authors.forename: Charlotte M
authors.initials: CM
authors.lastname: Flynn
authors.affiliation: Sir William Dunn School of Pathology, South Parks Road, Oxford OX1 3RE, UK.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Künzel
authors.affiliation: Institute of Biochemistry, Kiel University, Olshausenstr 40, 24098 Kiel, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Garbers
authors.affiliation: Sir William Dunn School of Pathology, South Parks Road, Oxford OX1 3RE, UK.
authors.forename: Matthew
authors.initials: M
authors.lastname: Freeman
authors.affiliation: Institute of Biochemistry, Kiel University, Olshausenstr 40, 24098 Kiel, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Grötzinger
authors.affiliation: Institute of Biochemistry, Kiel University, Olshausenstr 40, 24098 Kiel, Germany.|Sir William Dunn School of Pathology, South Parks Road, Oxford OX1 3RE, UK.
authors.forename: Stefan
authors.initials: S
authors.lastname: Düsterhöft
mesh_terms: D000072198:ADAM17 Protein / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D022163:Clathrin-Coated Vesicles / Q000378:metabolism*; D015536:Down-Regulation; D004789:Enzyme Activation / Q000187:drug effects; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D010703:Phorbol Esters / Q000494:pharmacology*; D011493:Protein Kinase C / Q000378:metabolism; D059748:Proteolysis / Q000187:drug effects; D015398:Signal Transduction
chemical_list: D010703:Phorbol Esters; C032974:12-O-retinoylphorbol-13-acetate; D011493:Protein Kinase C; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human