Background: Intrinsically disordered proteins are common regulators of protein-protein interactions, but little is known about their mechanisms of interaction. Results: Two intrinsically disordered protein domains, from ACTR and CREB-binding protein, interact through rapid association and slow conformational changes. Conclusion: Electrostatics governs the fast association, but the overall reaction is multistep. Significance: The slow conformational search may be common among intrinsically disordered proteins with many binding partners.

Epicardium-derived cells (EPDCs) play a fundamental role in embryonic cardiac development and are reactivated in the adult heart in response to myocardial infarction (MI). In this study, EPDCs from post-MI rat hearts highly expressed the ectoenzyme CD73 and secreted the profibrotic matricellular protein tenascin-C (TNC). CD73 on EPDCs extensively generated adenosine from both extracellular ATP and NAD. This in turn stimulated the release of additional nucleotides from a Brefeldin A-sensitive intracellular pool adenosine-AR signaling, forming a positive-feedback loop. AR activation, in addition, strongly promoted the release of major regulatory cytokines, such as IL-6, IL-11, and VEGF. TNC was found to stimulate EPDC migration and, together with ATP-P2XR signaling, to activate inflammasomes in EPDCs TLR4. Our results demonstrate that EPDCs are an important source of various proinflammatory factors in the post-MI heart controlled by purinergic and TNC signaling.-Hesse, J., Leberling, S., Boden, E., Friebe, D., Schmidt, T., Ding, Z., Dieterich, P., Deussen, A., Roderigo, C., Rose, C. R., Floss, D. M., Scheller, J., Schrader, J. CD73-derived adenosine and tenascin-C control cytokine production by epicardium-derived cells formed after myocardial infarction.

The biological function of adherent cell populations strongly depends on the physical and biochemical properties of extracellular matrix molecules. Therefore, numerous biocompatible cell carriers have been developed to specifically influence cell attachment, proliferation, cellular differentiation, and tissue formation for diverse cell culture applications and cell-based therapies. In the present study, we evaluated the mechanical and the cell biological properties of a novel, thin, and planar collagen scaffold. The cell carrier is based on fibrillar bovine collagen type I and exhibits a low material thickness coupled with a high mechanical stability as measured by tensile tests. The influence of this new biomaterial on cell viability, proliferation, and cell differentiation was analyzed using 5-bromo-2-deoxyuridine (BrdU) proliferation assay, immunocytochemistry, water-soluble tetrazolium salt-1 assay (WST-1), live cell imaging, and electron microscopy. Cell culture experiments with the human osteosarcoma cell line Saos-2, human mesenchymal stem cells, and rodent cardiomyocytes demonstrated the in vitro biocompatibility of this chemically noncrosslinked scaffold. Both the mechanical characteristics and the in vitro biocompatibility of this collagen type I carrier facilitate the engineering of thin transferable tissue constructs and offer new possibilities in the fields of cell culture techniques, tissue engineering, and regenerative medicine.

The demand for scaffolds comprised of natural materials such as collagen has increased in recent years. However, many scaffolds rely on chemical or physical modifications in order to comply with the necessary requirements for biomedical engineering. We evaluated the in vivo biocompatibility and biodegradation of a novel, thin, mechanically stable, and chemically non-crosslinked collagen cell carrier (CCC). CCC was implanted subcutaneously into 25 adult Lewis rats and biopsies were taken on days 7, 14, 21, 42, and 84 after surgery. For histological analysis, paraffin sections of implanted skin were immunolabeled for CD68 and stained by hematoxylin-eosin and Masson-Goldner's trichrome method. Macroscopic analysis of skin surface during wound healing process showed a normal physiological reaction. Biodegradation of CCC was completed 42 days after subcutaneous implantation. Histological evaluation revealed no evidence of encapsulation, scar formation, or long-term vascularization and inflammation. The collagen type I based biomaterial demonstrated a high in vivo biocompatibility, low irritability, complete resorption, and replacement by autologous tissue. The in vivo biocompatibility and degradation behavior encourage for further evaluation of CCC in surgical applications and regenerative medicine.

NON-PHOTOTROPIC HYPOCOTYL 3 (NPH3) is a key component of the auxin-dependent plant phototropic growth response. We report that NPH3 directly binds polyacidic phospholipids, required for plasma membrane association in darkness. We further demonstrate that blue light induces an immediate phosphorylation of a C-terminal 14-3-3 binding motif in NPH3. Subsequent association of 14-3-3 proteins is causal for the light-induced release of NPH3 from the membrane and accompanied by NPH3 dephosphorylation. In the cytosol, NPH3 dynamically transitions into membraneless condensate-like structures. The dephosphorylated state of the 14-3-3 binding site and NPH3 membrane recruitment are recoverable in darkness. NPH3 variants that constitutively localize either to the membrane or to condensates are non-functional, revealing a fundamental role of the 14-3-3 mediated dynamic change in NPH3 localization for auxin-dependent phototropism. This regulatory mechanism might be of general nature, given that several members of the NPH3-like family interact with 14-3-3 via a C-terminal motif.