title: Synthetic enzyme-substrate tethering obviates the Tolloid-ECM interaction during Drosophila BMP gradient formation.
abstract: Members of the Tolloid family of metalloproteinases liberate BMPs from inhibitory complexes to regulate BMP gradient formation during embryonic dorsal-ventral axis patterning. Here, we determine mechanistically how Tolloid activity is regulated by its non-catalytic CUB domains in the Drosophila embryo. We show that Tolloid, via its N-terminal CUB domains, interacts with Collagen IV, which enhances Tolloid activity towards its substrate Sog, and facilitates Tsg-dependent stimulation of cleavage. In contrast, the two most C-terminal Tld CUB domains mediate Sog interaction to facilitate its processing as, based on our structural data, Tolloid curvature positions bound Sog in proximity to the protease domain. Having ascribed functions to the Tolloid non-catalytic domains, we recapitulate embryonic BMP gradient formation in their absence, by artificially tethering the Tld protease domain to Sog. Our studies highlight how the bipartite function of Tolloid CUB domains, in substrate and ECM interactions, fine-tune protease activity to a particular developmental context.
authors: Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.
authors: Jennifer
authors: J
authors: Winstanley
authors: Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.
authors: Annick
authors: A
authors: Sawala
authors: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.
authors: Clair
authors: C
authors: Baldock
authors: Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.
authors: Hilary L
authors: HL
authors: Ashe
mesh_terms: D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D020134:Catalytic Domain; D024141:Collagen Type IV / Q000378:metabolism; D029721:Drosophila Proteins / Q000737:chemistry / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D008958:Models, Molecular; D050505:Mutant Proteins / Q000378:metabolism; D017354:Point Mutation; D011485:Protein Binding; D015202:Protein Engineering; D013379:Substrate Specificity; D055394:Tolloid-Like Metalloproteinases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D019485:Bone Morphogenetic Proteins; D024141:Collagen Type IV; D029721:Drosophila Proteins; D050505:Mutant Proteins; D055394:Tolloid-Like Metalloproteinases; C071471:tld protein, Drosophila
keywords: BMP; Collagen IV; D. melanogaster; Sog/Chordin; Tolloid; cell biology; developmental biology; gradient formation; stem cells
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