Wnt/-catenin signaling controls cell growth during development, and its misregulation in adults can cause human diseases. LRP6, the essential co-receptor for the Wnt pathway, consists of four -propeller domains flanked by epidermal growth factor repeats in its extracellular region. To understand the maturation and ligand-binding properties of individual BP domains, we generated soluble receptor consisting of individual BPs, as well as combinations of these domains. We show that BP1, BP2, and BP4 each can be folded and secreted, and their secretion was enhanced by co-expression of Mesd, a molecular chaperone essential for LRP6 folding and maturation. BP3 is not secreted when expressed on its own or in combination with BP2 or BP1 and 2 (BP12); however, folding and secretion of BP3 is vastly enhanced when expressed together with BP4. Similar cooperative folding and maturation was observed between BP1 and BP2. These results suggest that BP1 forms a functional folding unit with BP2, whereas BP3 folds together with BP4. Using these BP constructs, we also found that BP12 and BP34 constitute independent ligand-binding domains capable of binding Wnt3a, Dkk1, and Mesd. The ability of Mesd to block the binding of both Wnt3a and Dkk1 to LRP6 enables this specialized chaperone to function as a Wnt signaling modulator. Together, our studies reveal unique properties of the LRP6 BP domains and provide novel tools to understand LRP6 function in ligand binding and Wnt signaling. Our results also support the development of soluble LRP6 receptors and Mesd as potential therapeutic molecules that target Wnt signaling.
Indoor food production in vertical hydroponic systems can be done year-round in any climate, has less negative environmental impacts compared to industrial agriculture and is a way of increasing the nutritional value of crops. Indoor hydroponic systems make it easy to control the precision of additives and amendments, such as fertilizers and vermicompost tea, to nutrient solution reservoirs. Vermicompost tea is known to contain beneficial microorganisms and can help increase the biomass, nutrient density, and overall health of the plants. Microorganisms and plants have co-evolved and is an essential relationship that deserves recognition and further research. The goal of the research presented in this paper was to better understand if vermicompost tea and its associated microorganisms, such as bacteria, fungi, nematodes, and protozoa, would increase biomass and nutrient density of Purple Lady Bok Choy (Brassica rapa var. chinensis) in vertical hydroponic systems. Statistical analyses were performed to compare the biomass and macro- and micronutrients of three different treatments to one another and a control. There was a significant difference between the mean leaf and root biomass among varying concentrations of vermicompost tea solutions and added inorganic hydroponic fertilizer. The nutrient density for both macro- and micronutrients also differed significantly due to different concentrations of vermicompost tea, suggesting that beneficial microbes may help plants uptake and absorb nutrients in a more efficient manner depending on the concentration of vermicompost tea and hydroponic fertilizer.
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