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Summary: Inorganic polyphosphates (polyPs) are linear polymers of orthophosphate units linked by phosphoanhydride bonds. Polyphosphates represent important stores of phosphate and energy, and are abundant in many pro‐ and eukaryotic organisms. In plants, the existence of polyPs has been established using microscopy and biochemical extraction methods that are now known to produce artifacts. Here we use a polyP‐specific dye and a polyP‐binding domain to detect polyPs in plant and algal cells. To develop the staining protocol, we induced polyP granules in Nicotiana benthamiana and Arabidopsis cells by heterologous expression of Escherichia coli polyphosphate kinase 1 (PPK1). Over‐expression of PPK1 but not of a catalytically impaired version of the enzyme leads to severe growth phenotypes, suggesting that ATP‐dependent synthesis and accumulation of polyPs in the plant cytosol is toxic. We next crossed stable PPK1‐expressing Arabidopsis lines with plants expressing the polyP‐binding domain of E. coli exopolyphosphatase (PPX1c), which co‐localized with PPK1‐generated polyP granules. These granules were stained by the polyP‐specific dye JC‐D7 and appeared as electron‐dense structures in transmission electron microscopy sections. Using the polyP staining protocol derived from these experiments, we screened for polyP stores in different organs and tissues of both mono‐ and dicotyledonous plants. While we could not detect polyP granules in higher plants, we could visualize the polyP‐rich acidocalcisomes in the green alga Chlamydomonas reinhardtii. Significance Statement: A chemical dye and an inorganic polyphosphate‐binding domain are shown to specifically label inorganic polyphosphate granules in transgenic Arabidopsis lines and Chlamydomonas acidocalcisomes. Using these tools we show that, in contrast to many prokaryotic and eukaryotic organisms, higher plants do not seem to contain large stores of inorganic polyphosphate. [ABSTRACT FROM AUTHOR]

Hydrogen sulfi de (H2S) has been recognized as one of most important gaseous signaling molecules mediated by a variety of physiological and pathological processes. Yet, its functions remain largely elusive due to the lack of potent monitoring methods. Hereby this issue is addressed with a powerful new platform--dye-assembled upconversion nanoparticles (UCNPs). A series of chromophores with different absorption bands and fast responses towards H2S is combined with UCNPs and results in a library of H2S sensors with responsive emission signals ranging from the visible to the near-infrared (NIR) region. These nanoprobes demonstrate highly selective and rapid responses to H2S in vitro and in cells. Furthermore, H2S levels in blood can be detected using the developed nanoprobes. Hence the reported H2S sensing platform can serve as a powerful diagnostic tool to research H2S functions and to investigate H2S-related diseases. [ABSTRACT FROM AUTHOR]

As most of the available depigmenting agents exhibit only modest activity and some exhibit toxicities that lead to adverse side effects after long-term usage, there remains a need for novel depigmenting agents. Chemical genetic screening was performed on cultured melanocytes to identify novel depigmenting compounds. By screening a tagged-triazine library, we identified four compounds, TGH11, TGD10, TGD39 and TGJ29, as potent pigmentation inhibitors with IC50 values in the range of 10 μM. These newly identified depigmenting compounds were found to function as reversible inhibitors of tyrosinase, the key enzyme involved in melanin synthesis. Tyrosinase was further confirmed as the cellular target of these compounds by affinity chromatography. Kinetic data suggest that all four compounds act as competitive inhibitors of tyrosinase, most likely competing with l-3,4-dihydroxyphenylalanine ( l-DOPA) for binding to the DOPA-binding site of the enzyme. No effect on levels of tyrosinase protein, processing or trafficking was observed upon treatment of melanocytes with these compounds. Cytotoxicity was not observed with these compounds at concentrations up to 20 μM. Our data suggest that TGH11, TGD10, TGD39 and TGJ29 are novel potent tyrosinase inhibitors with potential beneficial effects in the treatment of cutaneous hyperpigmentation. [ABSTRACT FROM AUTHOR]

Examines the cloning of the recombinant p42[sup IP4], a membrane-associated and cytosolic inositol tetrakisphosphate receptor protein. Use of protein-lipid vesicle binding assay; Analysis of the protein using Western blotting; Identification of inositol phosphates and lipids.