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Summary: Transient receptor potential melastatin 2 (TRPM2), a Ca2+-permeable cation channel, is gated by intracellular adenosine diphosphate ribose (ADPR), Ca2+, warm temperature, and oxidative stress. It is critically involved in physiological and pathological processes ranging from inflammation to stroke to neurodegeneration. At present, the channel’s gating and ion permeation mechanisms, such as the location and identity of the selectivity filter, remain ambiguous. Here, we report the cryo-electron microscopy (cryo-EM) structure of human TRPM2 in nanodisc in the ligand-free state. Cryo-EM map-guided computational modeling and patch-clamp recording further identify a quadruple-residue motif as the ion selectivity filter, which adopts a restrictive conformation in the closed state and acts as a gate, profoundly contrasting with its widely open conformation in the Nematostella vectensis TRPM2. Our study reveals the gating of human TRPM2 by the filter and demonstrates the feasibility of using cryo-EM in conjunction with computational modeling and functional studies to garner structural information for intrinsically dynamic but functionally important domains.

The transformation of a ciliate into cyst is an advance strategy against an adverse situation. However, the molecular mechanism for the encystation of free-living ciliates is poorly understood. A large-scale identification of the encystment-related proteins and genes in ciliate would provide us with deeper insights into the molecular mechanisms for the encystations of ciliate. We identified the encystment-related proteins and genes in Pseudourostyla cristata with shotgun LC-MS/MS and scale qRT-PCR, respectively, in this report. A total of 668 proteins were detected in the resting cysts, 102 of these proteins were high credible proteins, whereas 88 high credible proteins of the 724 total proteins were found in the vegetative cells. Compared with the vegetative cell, 6 specific proteins were found in the resting cyst. However, the majority of high credible proteins in the resting cyst and the vegetative cell were co-expressed. We compared 47 genes of the co-expressed proteins with known functions in both the cyst and the vegetative cell using scale qRT-PCR. Twenty-seven of 47 genes were differentially expressed in the cyst compared with the vegetative cell. In our identifications, many uncharacterized proteins were also found. These results will help reveal the molecular mechanism for the formation of cyst in ciliates.

The transformation of a ciliate into cyst is an advance strategy against an adverse situation. However, the molecular mechanism for the encystation of free-living ciliates is poorly understood. A large-scale identification of the encystment-related proteins and genes in ciliate would provide us with deeper insights into the molecular mechanisms for the encystations of ciliate. We identified the encystment-related proteins and genes in Pseudourostyla cristata with shotgun LC-MS/MS and scale qRT-PCR, respectively, in this report. A total of 668 proteins were detected in the resting cysts, 102 of these proteins were high credible proteins, whereas 88 high credible proteins of the 724 total proteins were found in the vegetative cells. Compared with the vegetative cell, 6 specific proteins were found in the resting cyst. However, the majority of high credible proteins in the resting cyst and the vegetative cell were co-expressed. We compared 47 genes of the co-expressed proteins with known functions in both the cyst and the vegetative cell using scale qRT-PCR. Twenty-seven of 47 genes were differentially expressed in the cyst compared with the vegetative cell. In our identifications, many uncharacterized proteins were also found. These results will help reveal the molecular mechanism for the formation of cyst in ciliates.

Wild bananas are abundant in tropical areas and many ecologists have observed that the succession process is quicker following increased disturbance. This study was conducted to analyze animal-seed interactions and their effects on the seed fate of a wild banana species (Musa acuminata) in tropical southern Yunnan (China) through experiments considering spatial (site and habitat) and temporal (seasons) variation. The largest proportion of fruits (81%) was removed by frugivorous seed dispersers, especially by bats at nighttime. Only 13% of the fruits were removed by climbing seed predators (different species of rats). In the exclosure treatment, rodents accounted for a significantly higher total artificially exposed seed removal number than ants, but with spatial and temporal differences. The highest seed predation rate by rodents (70%) was found in forest with wild banana stands, corresponding with the highest rodent diversity (species numbers and abundance) among the habitat types. In contrast, the seed removal number by ants (57%) was highest in the open land habitats, but there was no close correlation with ant diversity. Seed removal numbers by ants were significantly higher in the dry compared to the rainy season, but rodent activity showed no differences between seasons. The overall results suggest that the largest proportion of seeds produced by wild banana are primarily dispersed by bats. Primary seed dispersal by bats at nighttime is essential for wild banana seeds to escape seed predation.

Davood Nematollahi et al. have successfully electrosynthesised a series of benzofuran derivatives in the presence of nucleophile agents. Base on their experimental result, we analyzed the electron density of the intermediate during benzofuran derivatives synthesis and reached the same conclusions as that of Davood Nematollahi et al..Therefore, the method of electron density analysis is proven to be correct and expected to have a wide applications. But after we analyzed the electron density of 2-aldehyde-p-quinone(1a) and 3-carboxyl-o-quinone(7a), we have different opinions with theirs. The electron density of C-6 position of the 2-aldehyde-p-quinone lower than the that of C-5 position leads to the final product 6 when electrochemical oxidation of the 2,5-dihydroxybenzaldehyde. For 2,3-dihydroxybenzoic acid case, since the competition between the carboxyl of the C-2 position and the carboxyl of the C-1’ position in the 3-carboxyl-o-quinone and the hydrogen transferred to the carboxyl of the C-1’ position, there will be a rearrangement before the next electrochemical oxidation and the final product 21 will be formed.