1. Incorporation of Pentraxin 3 into Hyaluronan Matrices Is Tightly Regulated and Promotes Matrix Cross-linking
- Author
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Baranova, NS, Inforzato, A, Briggs, DC, Tilakaratna, V, Enghild, JJ, Thakar, D, Milner, CM, Day, AJ, and Richter, RP
- Subjects
Quartz Crystal Microbalance ,Biophysics ,Glycobiology ,Glycobiology and Extracellular Matrices ,Supramolecular Structure ,Extracellular Matrix Protein ,Hyaluronate ,Reflection Interference-Contrast Microscopy ,Spectroscopic Ellipsometry ,Analytical Chemistry ,Cell Line ,Extracellular Matrix ,Serum Amyloid P-Component ,C-Reactive Protein ,Drosophila melanogaster ,Ovarian Follicle ,Alpha-Globulins ,Animals ,Humans ,Female ,Hyaluronic Acid ,Carbohydrate-binding Protein ,Cell Adhesion Molecules ,Protein Binding - Abstract
Background: The proteins pentraxin 3 (PTX3) and TNF-stimulated gene-6 (TSG-6) and the proteoglycan inter-α-inhibitor (IαI) are known to be involved in the stabilization of hyaluronan (HA)-rich extracellular matrices. Results: PTX3 incorporation into HA matrices is tightly regulated. Conclusion: PTX3, TSG-6, and IαI are sufficient to cross-link HA matrices. Significance: The results provide mechanistic insights into the regulation of HA-protein interactions., Mammalian oocytes are surrounded by a highly hydrated hyaluronan (HA)-rich extracellular matrix with embedded cumulus cells, forming the cumulus cell·oocyte complex (COC) matrix. The correct assembly, stability, and mechanical properties of this matrix, which are crucial for successful ovulation, transport of the COC to the oviduct, and its fertilization, depend on the interaction between HA and specific HA-organizing proteins. Although the proteins inter-α-inhibitor (IαI), pentraxin 3 (PTX3), and TNF-stimulated gene-6 (TSG-6) have been identified as being critical for COC matrix formation, its supramolecular organization and the molecular mechanism of COC matrix stabilization remain unknown. Here we used films of end-grafted HA as a model system to investigate the molecular interactions involved in the formation and stabilization of HA matrices containing TSG-6, IαI, and PTX3. We found that PTX3 binds neither to HA alone nor to HA films containing TSG-6. This long pentraxin also failed to bind to products of the interaction between IαI, TSG-6, and HA, among which are the covalent heavy chain (HC)·HA and HC·TSG-6 complexes, despite the fact that both IαI and TSG-6 are ligands of PTX3. Interestingly, prior encounter with IαI was required for effective incorporation of PTX3 into TSG-6-loaded HA films. Moreover, we demonstrated that this ternary protein mixture made of IαI, PTX3, and TSG-6 is sufficient to promote formation of a stable (i.e. cross-linked) yet highly hydrated HA matrix. We propose that this mechanism is essential for correct assembly of the COC matrix and may also have general implications in other inflammatory processes that are associated with HA cross-linking. more...
- Published
- 2014
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