ADAMTS10 Protein Interacts with Fibrillin-1 and Promotes Its Deposition in Extracellular Matrix of Cultured Fibroblasts

Abstract

Autosomal recessive and autosomal dominant forms of WeillMarchesani syndrome, an inherited connective tissue disorder, are caused by mutations in ADAMTS10 (encoding a secreted metalloprotease) and FBN1 (encoding fibrillin-1, which forms tissue microfibrils), respectively, yet they are clinically indistinguishable. This genetic connection prompted investigation of a potential functional relationship between ADAMTS10 and fibrillin-1. Specifically, fibrillin-1 was investigated as a potential ADAMTS10 binding partner and substrate, and the role of ADAMTS10 in influencing microfibril biogenesis was addressed. Using ligand affinity blotting and surface plasmon resonance, recombinant ADAMTS10 was found to bind to fibrillin-1 with a high degree of specificity and with high affinity. Two sites of ADAMTS10 binding to fibrillin-1 were identified, one toward the N terminus and another in the C-terminal half of fibrillin-1. Confocal microscopy and immunoelectron microscopy localized ADAMTS10 to fibrillin-1-containing microfibrils in human tissues. Furin-activated ADAMTS10 could cleave fibrillin-1, but innate resistance of ADAMTS10 zymogen to propeptide excision by furin was observed, suggesting that, unless activated, ADAMTS10 is an inefficient fibrillinase. To investigate the role of ADAMTS10 in microfibril biogenesis, fetal bovine nuchal ligament cells were cultured in the presence or absence of ADAMTS10. Exogenously added ADAMTS10 led to accelerated fibrillin-1 microfibril biogenesis. Conversely, fibroblasts obtained from a Weill-Marchesani syndrome patient with ADAMTS10 mutations deposited fibrillin-1 microfibrils sparsely compared with unaffected control cells. Taken together, these findings suggest that ADAMTS10 participates in microfibril biogenesis rather than in fibrillin-1 turnover.Inherited connective tissue disorders that result from mutations of genes encoding extracellular matrix (ECM) 3 components or their modifying enzymes are potentially insightful in identifying structural and regulatory roles of ECM constituents or their supramolecular assemblies. A classic example of such an ECM component is fibrillin-1, a large, secreted glycoprotein, which is a major component of 8 -12-nm-diameter tissue microfibrils (1). Bundles of microfibrils, containing fibrillin-1, fibrillin-2, and fibrillin-3 (in humans), are ubiquitous in the body but are especially abundant in tissues containing elastin (2). The structural role of microfibrils includes maintenance of the ocular lens in its appropriate position in the eye via the zonule of Zinn (the suspensory ligament of the lens), which comprises primarily fibrillin-1 microfibrils (2, 3). Inadequacy of the zonule or traumatic breakage of zonule fibers can result in subluxation or dislocation of the lens (ectopia lentis). Dominantly inherited FBN1 mutations lead to diverse connective tissue anomalies, the most common of which is Marfan syndrome (MFS) (4). Major features of MFS include musculoskeletal anomalies, aortic aneurysms, and ectopia lentis. Several features of MFS appear...

The Microfibril Hypothesis of Glaucoma: Implications for Treatment of Elevated Intraocular Pressure

“…6 Although Weill-Marchesani patients are opposite to Marfan patients in outward appearance, the syndromes share a common molecular mechanism, which is defective fibrillin-1 microfibrils. 33,73,77 Further supporting a role for ADAMTS10 in microfibril function, co-localization of ADAMTS10 and fibrillin-1 has been shown by immunohistochemistry of human skin 33,73 and ADAMTS10 has been shown to promote microfibril formation in cell culture. 73 Specific and high-affinity binding of ADAMTS10 to fibrillin-1 has been demonstrated by affinity blotting assays, affinity pull-down assays, and surface plasmon resonance.…”

“…33,73,77 Further supporting a role for ADAMTS10 in microfibril function, co-localization of ADAMTS10 and fibrillin-1 has been shown by immunohistochemistry of human skin 33,73 and ADAMTS10 has been shown to promote microfibril formation in cell culture. 73 Specific and high-affinity binding of ADAMTS10 to fibrillin-1 has been demonstrated by affinity blotting assays, affinity pull-down assays, and surface plasmon resonance. 33,73 The binding site on fibrillin-1 for ADAMTS10 overlaps with the binding site for other ADAMTS proteins, 33 suggesting competition for the fibrillin-1 binding site or complex formation with other microfibril-associated proteins.…”

“…1,71 ADAMTS10 is a secreted matrix metalloproteinase that can cleave fibrillin-1. 72,73 Although its exact function is not known, a role for ADAMTS10 in microfibril structure and function was first suggested by the finding that Weill-Marchesani syndrome can be caused either by recessive ADAMTS10 mutations 74 or dominant mutations in FBN1. 75 Clinically, the dominant and recessive forms of Weill-Marchesani are indistinguishable, suggesting common functional roles for ADAMTS10 and FBN1.…”

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“…MMPs degrade collagen and elastin in ECM and degradation of collagen is one of the most important events in invasion and metastasis. 29 32 Kutz et al 33 showed that there was a direct interaction between ADAMTS10 and fibrillin-1 in ECM. Like other ADAMTS, ADAMTS10 mutations may change biomechanical features of ECM in animals.…”

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“…Interestingly, there are some reports regarding noncatalytic activities of ADAMTSs. Mammalian ADAMTS10 participates in microfibril biogenesis through its binding to fibrillin-1, in which the catalytic activity of ADAMTS10 appears not to be involved (Kutz et al 2011). ADAMTS4 binds and colocalizes with fibronectin at the cell surface, where fibronectin can inhibit ADAMTS4 activity as an aggrecanase (Hashimoto et al 2004).…”