Protein Nanobarrel for Integrating Chlorophyll a Molecules and Its Photochemical Performance.

Taking inspiration from biology's effectiveness in nanoscale organization of chlorophylls for photosynthesis, we describe here a design for chlorophyll-protein conjugates that exploits the central hydrophobic cavity of GroEL protein nanobarrel as a binding pocket for chlorophyll. We found water-soluble conjugates of chlorophyll with GroEL could be easily generated via detergent dialysis. The number of chlorophyll units bound to GroEL is tunable by varying the equilibrium concentration of chlorophyll during dialysis. Meanwhile, it is shown that an increase in the entrapped chlorophyll amount leads to an improvement of chlorophyll-GroEL photostability. Using methyl viologen as an electron acceptor, we demonstrate that chlorophyll-GroEL has photoreduction activity, which is also switchable in on/off illumination mode. Finally, it is shown that chlorophyll-GroEL-sensitized solar cells have good photoelectric properties, yielding a high photoelectric conversion efficiency of ∼0.9%. The current strategy may be adopted for integrating other photosensitizing dyes or for other photocatalytic reactions.

Down-regulation of PCK2 inhibits the invasion and metastasis of laryngeal carcinoma cells.

Laryngeal carcinoma is one of the common malignancies of head and neck. However, the pathogenesis of laryngeal cancer has been not completely clear. To identify the effects of hypoxia on the invasion, metastasis, and metabolism of laryngeal carcinoma, iTRAQ-labeling-with-LC-MS/MS analysis was performed to identify differentially expressed proteins of the SCC10A cells under hypoxia and normoxia, while metabolites were examined by metabolic profiling. 155 proteins and 180 metabolites were identified and the PCK2 protein was selected for validation by Western Blotting. Immunohistochemistry (IHC) was performed to analyze the expression of PCK2 in formalin-fixed paraffin-embedded (FFPE) tissue sections, including laryngeal squamous cell carcinoma tissues from various stages. Collectively, we report that down-regulation of PCK2 inhibits the invasion, migration, and proliferation of laryngeal cancer under hypoxia and down-regulation of PCK2 may be used as a new strategy for laryngeal cancer therapy.

Characterization of the complete mitochondrial genome of yellow stripe mutant in onion (Allium cepa L.).

The complete onion of yellow stripe mutant tissues mitochondrial genome was determined and analyzed in this study. The complete sequence length is 324,182 bp, including 37 CDS, 13 tRNA, and G + C content percentage of 45. Compared with the normal onions, 556 SNPs, 279 small indel numbers were detected, and the mutations of 14 SNPs and two small indels in the exon region affected the translation of proteins. Phylogenetic tree analysis showed that yellow stripe mutant onion is closely clustered with other Allium cepa L. The mutant of onion plays an important role in developing molecular marker-assisted breeding.

Ergosterol peroxide inhibits ovarian cancer cell growth through multiple pathways.

Ergosterol peroxide (EP), a sterol derived from medicinal mushrooms, has been reported to exert antitumor activity in several tumor types. However, the role of EP toward ovarian cancer cells has not been investigated. In this study, we analyzed the cytotoxicity of EP in various cell lines representing high-grade serous ovarian cancer and low-grade serous ovarian cancer, respectively. Although EP showed no significant inhibition of the viability of normal ovarian surface epithelial cells, it impaired the proliferation and invasion capacities of tumor cells in a dose-dependent manner. We further figured out key modulators involved in its antitumor effects by quantitative reverse transcription polymerase chain reaction, ELISA, and Western blot. The nuclear ß-catenin was down-regulated upon EP treatment, subsequently reducing the Cyclin D1 and c-Myc expression levels. Meanwhile, the protein level of protein tyrosine phosphatase SHP2 was up-regulated in EP treated cells, whereas Src kinase activity was inhibited. Both activation of SHP2 phosphatase and inhibition of Src kinase decreased the phosphorylation level of transducer and activator of STAT3 protein, which was implicated in oncogenesis. On the other hand, EP remarkably inhibited the expression and secretion of VEGF-C, implying its involvement in counteracting tumor angiogenesis. Moreover, EP treatment showed comparable cytotoxic effect with ß-catenin knock-down or STAT3 inhibition. Taken together, our results demonstrated that EP showed antitumor effects toward ovarian cancer cells through both ß-catenin and STAT3 signaling pathways, making it a promising candidate for drug development.

Chaperonin-Nanocaged Hemin as an Artificial Metalloenzyme for Oxidation Catalysis.

Taking inspiration from biology's effectiveness in functionalizing protein-based nanocages for chemical processes, we describe here a rational design of an artificial metalloenzyme for oxidations with the bacterial chaperonin GroEL, a nanocage for protein folding in nature, by supramolecular anchoring of catalytically active hemin in its hydrophobic central cavity. The promiscuity of the chaperonin cavity is an essential element of this design, which can mimic the hydrophobic binding pocket in natural metalloenzymes to accept cofactor and substrate without requiring specific ligand-protein interactions. The success of this approach is manifested in the efficient loading of multiple monomeric hemin cofactors to the GroEL cavity by detergent dialysis and good catalytic oxidation properties of the resulting biohybrid in tandem with those of the clean oxidant of H2O2. Investigation of the mechanism of hemin-GroEL-catalyzed oxidation of two-model substrates reveals that the kinetic behavior of the complex follows a ping-pong mechanism in both cases. Through comparison with horseradish peroxidase, the oxidative activity and stability of hemin-GroEL were observed to be similar to those found in natural peroxidases. Adenosine 5'-triphosphate (ATP)-regulated partial dissociation of the biohybrid, as assessed by the reduction of its catalytic activity with the addition of the nucleotide, raises the prospect that ATP may be used to recycle the chaperonin scaffold. Moreover, hemin-GroEL can be applied to the chromogenic detection of H2O2, which (or peroxide in general) is commonly contained in industrial wastes. Considering the rich chemistry of free metalloporphyrins and the ease of production of GroEL and its supramolecular complex with hemin, this work should seed the creation of many new artificial metalloenzymes with diverse reactivities.