title: C-terminal propeptide is required for fibrillin-1 secretion and blocks premature assembly through linkage to domains cbEGF41-43.
abstract: Fibrillin microfibrils are 10-12 nm diameter, extracellular matrix assemblies that provide dynamic tissues of metazoan species with many of their biomechanical properties as well as sequestering growth factors and cytokines. Assembly of fibrillin monomers into microfibrils is thought to occur at the cell surface, with initial steps including proprotein processing, multimerization driven by the C terminus, and the head-to-tail alignment of adjacent molecules. At present the mechanisms that regulate microfibril assembly are still to be elucidated. We have used structure-informed protein engineering to create a recombinant, GFP-tagged version of fibrillin-1 (GFP-Fbn) to study this process. Using HEK293T cells transiently transfected with GFP-Fbn constructs, we show that (i) the C-terminal propeptide is an essential requirement for the secretion of full-length fibrillin-1 from cells; (ii) failure to cleave off the C-terminal propeptide blocks the assembly of fibrillin-1 into microfibrils produced by dermal fibroblasts; and (iii) the requirement of the propeptide for secretion is linked to the presence of domains cbEGF41-43, because either deletion or exchange of domains in this region leads to cellular retention. Collectively, these data suggest a mechanism in which the propeptide blocks a key site at the C terminus to prevent premature microfibril assembly.
authors: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom sacha.jensen@bioch.ox.ac.uk penny.handford@bioch.ox.ac.uk.
authors: Sacha A
authors: SA
authors: Jensen
authors: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.
authors: Georgia
authors: G
authors: Aspinall
authors: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom sacha.jensen@bioch.ox.ac.uk penny.handford@bioch.ox.ac.uk.
authors: Penny A
authors: PA
authors: Handford
mesh_terms: D000818:Animals; D020405:Dermis / Q000166:cytology / Q000378:metabolism*; D000071838:Fibrillin-1; D000071837:Fibrillins; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D020894:Microfibrils / Q000235:genetics / Q000378:metabolism*; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D054392:Adipokines
chemical_list: D000071838:Fibrillin-1; D000071837:Fibrillins; D008840:Microfilament Proteins; D011993:Recombinant Fusion Proteins; C000605722:FBN1 protein, human; D054392:Adipokines