In addition to its role in forming vesicles from the endoplasmic reticulum (ER), the coat protein complex II (COPII) is also responsible for selecting specific cargo proteins to be packaged into COPII transport vesicles. Comparison of COPII vesicle formation in mammalian systems and in yeast suggested that the former uses more elaborate mechanisms for cargo recognition, presumably to cope with a significantly expanded repertoire of cargo that transits the secretory pathway. Using proTGFα, the transmembrane precursor of transforming growth factor α (TGFα), as a model cargo protein, we demonstrate in cell-free assays that at least one auxiliary cytosolic factor is specifically required for the efficient packaging of proTGFα into COPII vesicles. Using a knockout HeLa cell line generated by CRISPR/Cas9, we provide functional evidence showing that a transmembrane protein, Cornichon-1 (CNIH), acts as a cargo receptor of proTGFα. We show that both CNIH and the auxiliary cytosolic factor(s) are required for efficient recruitment of proTGFα to the COPII coat in vitro. Moreover, we provide evidence that the recruitment of cargo protein by the COPII coat precedes and may be distinct from subsequent cargo packaging into COPII vesicles.

In this study, we established a new methodology for preparing 5-nucleotidase (5-NT) with the aim of enhancing our understanding of its enzyme activity and laying a basis for regulating the content of umami-enhancing nucleotides in pork. 5-NT was prepared with Sephadex gel filtration and reverse-phase high-performance liquid chromatography, and its enzymatic properties and catalytic activity were evaluated. The results show that the molecular weight of the prepared 5-NT was 57 kDa, the optimal catalytic temperature was 40 °C, and the optimal pH was 8. Zn2+, and sucrose showed inhibitory effects on the activity of 5-NT, while K+, Na+, Ca2+, Mg2+, glucose, fructose, and trehalose promoted the activity of the studied compound. The prepared 5-NT exhibited higher catalytic activity and selectivity against IMP compared with its commercial counterpart, while its catalytic activity against XMP was not significant (p > 0.05). In brief, we established a new methodology for preparing 5-NT, enhancing our understanding of its enzyme activity and providing a solid basis for regulating the content of umami-enhancing nucleotides in pork through the control of endogenous 5-NT activity.

To extend the survival of crucian carp (Carassius auratus) during transportation, the effect of deep dormancy temperature (DDT) cultivation on the crucian carp was investigated by measuring the respiratory rate, survival time, and effect of cooling speed on the meat quality. The results of the respiratory rate and survival time indicated that the DDT of the crucian carp was 1.6 °C. The cooling speed had a significant (p < 0.05) influence on the quality of the crucian carp meat, with a faster cooling speed resulting in a lower pH, L* value, a* value, gumminess, springiness, cohesiveness, stickiness, chewiness, CMP, and UMP content for the crucian carp meat, thus resulting in a lower sensory score for the crucian carp meat. A possible reason for the decrease in the quality of the crucian carp meat is that the faster cooling speed led to a strong stress response and higher anaerobic metabolism in the crucian carp. This can be supported by the contents of the blood glucose and lactic acid in the crucian carp treated with higher cooling speed being significantly (p < 0.05) higher than that of the control. Combining all the results of the cooling speed on the eating quality of the crucian carp meat, a cooling speed of 2 °C/h followed by 1 °C/h was suggested for the survival of crucian carp in transit.

The East Asian summer monsoon (EASM) supplies vital rainfall for over one billion people. El Niño-Southern Oscillation (ENSO) markedly affects the EASM, but its impacts are more robust following El Niño than La Niña. Here, we show that this asymmetry arises from the asymmetry in ENSO evolution: though most El Niño events last for one year, La Niña events often persist for 2-3 years. In the summers between consecutive La Niña events, the concurrent La Niña opposes the delayed effect of the preceding winter La Niña on the EASM, causing a reduction in the magnitude and coherence of climate anomalies. Results from a large ensemble climate model experiment corroborate and strengthen the observational analysis with an order of magnitude increase in sample size. The apparent asymmetry in the impacts of the ENSO on the EASM can be reduced by considering the concurrent ENSO, in addition to the ENSO state in the preceding winter. This has important implications for seasonal climate forecasts.

Silver carp (Hypophthalmichthys molitrix) is normally cooked at a high temperature. In order to explore the effects of cooking methods on the protein nutritional values and volatile flavor substances of silver carp meat, the meat was fried, roasted and steamed, and its contents were determined in relation to moisture, protein, fat, ash, amino acids and volatile flavor substances. The results show that the moisture content of cooked silver carp can be ranked as follows: raw > steamed > roasted > fried. The protein content order of the four samples can be ranked: fried > roasted > raw > steamed. The ash and the fat contents of the four samples showed similar significant (p < 0.05) differences, in the order of fried > roasted > steamed > raw. The contents of all the amino acids in the fried meat were significantly (p < 0.05) higher than the contents in others, and the frying, roasting and steaming processes improved the umami taste (supported by the increased content of glutamate and aspartate) and the protein nutritional value (supported by the amino acid score, essential amino acid index and protein efficiency ratio) of the silver carp meat. A total of 51 compounds were detected in the fried, roasted and steamed meat. Frying and roasting reduced the fishy flavor of this meat. The odor activity value (OAV) indicated that methanethiol, 1-octen-3-ol and 2-heptanone were the key flavor chemicals in raw meat. Hexanal, propionaldehyde, octanal, nonanal, decanal, 1-octen-3-ol and 2-heptanone were key to the flavor of fried meat. Pentanal, decanal, methyl mercap-tan and 1-octen-3-ol contributed greatly to the overall flavor of the roasted meat. Nonanal, methylmercaptan and 1-octene-3-ol had strong effects on the flavor of the steamed silver carp meat.

2D lamellar membranes (2DLMs) are used for efficient desalination and nanofiltration. However, weak interactions between adjacent stacked nanosheets result in susceptibility to swelling that limits practical applicability. Inspired by the super adhesion of multi-point suction cups on octopus tentacles, a 2DLM is constructed from Ti₃ C₂ T_x MXene supported by the macrocyclic "multi-point" molecule cucurbit[5]uril (CB5) and demonstrated for nanofiltration of methyl blue (MB) and enrichment of uranyl carbonate. Experimental results and density functional theory calculations indicate that CB5 rivets to the surface of the nanoflakes through strong stable interactions between its multiple binding sites and surface hydroxyl functional groups on MXene nanosheets. This novel 2DLM exhibits excellent nanofiltration performance (69 L m^-2 h^-1 bar^-1 permeance with 93.6% rejection for MB) and can be recycled at least 30 times without significant degradation. The 2DLM exhibits excellent swelling resistance at high salinity, with a demonstration of selective enrichment of uranyl carbonate from artificial water and natural seawater. The results provide a new strategy for constructing highly stable 2DLMs with interlayer spacing controllable from sub-nano to nanometer scales, for size-selective sieving of molecules and ions, high-efficiency nanofiltration, and other applications.

To evaluate the storage stability and flavor changes of marinated pork treated with chili and pepper essential oils, the contents of total sulfhydryl, malondialdehyde, total volatile base nitrogen (TVBN), Ca²⁺ATPase activity, and total viable counts of marinated pork were determined. Further, the non-volatile (umami, numb, and spicy) and volatile flavor compounds of marinated pork were analyzed. Based on the results, the chili and pepper essential oils had limited effects on the storage stability of marinated pork. However, these essential oils could inhibit the oxidation of lipids and proteins and reduce the number of microorganisms and TVBN in marinated pork within 6 days. The non-volatile flavors of the marinated pork decreased as the refrigeration time increased. It was concluded that the decomposition of umami-enhancing nucleotides (GMP, IMP, XMP), the number of flavor substances (hydroxyl-α-sanshool, hydroxyl-β-sanshool), and spicy (capsaicin) tasting compounds caused the decrease in non-volatile flavors.

To characterize the quality of widely consumed pork in China, the chemical compositions and other indexes of five breeds of pork were compared. The results indicated that the moisture content, sensory flavor, and overall acceptability of Pipa pork (PPP) were superior to other breeds. The fat content and essential amino acids (EAA) of Yihao native pork (YNP) were significantly (p < 0.05) higher than in other breeds. The protein content, the total amount of amino acids, and perceptible flavor of Tibetan black pork (TBP) were higher than in other breeds. The protein nutrition profiles of manor black pork (MBP) and TBP were better than in others. The equivalent umami concentration (EUC) value of white pork (WP) was significantly (p < 0.05) higher than in others; however, the health risk of its fatty acid content was higher. There were unique protein and flavor chemicals in YNP, TBP, and PPP, which may be useful for distinguishing their authenticity.

The secretory pathway of eukaryotic cells packages cargo proteins into COPII-coated vesicles for transport from the endoplasmic reticulum (ER) to the Golgi. We now report that complete genetic deficiency for the COPII component SEC24A is compatible with normal survival and development in the mouse, despite the fundamental role of SEC24 in COPII vesicle formation and cargo recruitment. However, these animals exhibit markedly reduced plasma cholesterol, with mutations in Apoe and Ldlr epistatic to Sec24a, suggesting a receptor-mediated lipoprotein clearance mechanism. Consistent with these data, hepatic LDLR levels are up-regulated in SEC24A-deficient cells as a consequence of specific dependence of PCSK9, a negative regulator of LDLR, on SEC24A for efficient exit from the ER. Our findings also identify partial overlap in cargo selectivity between SEC24A and SEC24B, suggesting a previously unappreciated heterogeneity in the recruitment of secretory proteins to the COPII vesicles that extends to soluble as well as trans-membrane cargoes. DOI:http://dx.doi.org/10.7554/eLife.00444.001.

Despite that two-dimensional transition metal carbides and carbonitrides (MXenes) are burgeoning candidates for remediation of environmental pollutants, the construction of robust functionalized MXene nanosheets with a high affinity for target heavy metal ions and radionuclides remains a challenge. Here we report the successful placement of amidoxime chelating groups on Ti₃C₂T_x MXene surface by diazonium salt grafting. The introduction of amidoxime functional groups significantly enhances the selectivity of Ti₃C₂T_x nanosheets for uranyl ions and also greatly improves their stability in aqueous solution, enabling efficient, rapid, and recyclable uranium extraction from aqueous solutions containing competitive metal ions. Benefiting from the excellent conductivity of MXenes, the amidoxime functionalized Ti₃C₂T_x nanosheets show outstanding electrochemical performance such that when loaded on carbon cloth the application of an electric field increases the uranium adsorption capacity from 294 to 626 mg/g, outperforming all organic electrochemical sorption materials reported previously. The present work provides an effective strategy to functionalize MXene nanosheets with fundamental implications for the design of MXene-based selective electrosorption electrode materials.