Upregulation of Divalent Metal Transporter 1 (DMT1) Is Involved in Amyloid Precursor Protein Processing and Aβ Generation

The amyloid beta precursor protein (APP) and its pathogenic byproduct β-amyloid peptide (Aβ) play central roles in the pathogenesis of Alzheimer’s disease (AD). Reduction in

Full-length core sequence dependent complex-type glycosylation of hepatitis C virus E2 glycoprotein

AIM: To study HCV polyprotein processing is important for the understanding of the natural history of HCV and the design of vaccines against HCV. The purpose of this study is to investigate the affection of context sequences on hepatitis C virus (HCV) E2 processing. METHODS: HCV genes of different lengths were expressed and compared in vaccinia virus/T7 system with homologous patient serum S94 and mouse anti-serum M( E2116) raised against E.coli -derived E2 peptide, respectively.Deglycosylation analysis and GNA ( Galanthus nivalus ) lectin binding assay were performed to study the post-translational processing of the expressed products. RESULTS: E2 glycoproteins with different molecular weights (-75 kDa and -60 kDa) were detected using S94 and M( E2116), respectively. Deglycosylation analysis showed that this difference was mainly due to different glycosylation. Endo H resistance and its failure to bind to GNA lectin demonstrated that the higher molecular weight form (75 kDa) of E2 was complex-type glycosylated, which was readily recognized by homologous patient serum S94. Expression of complex-type glycosylated E2 could not be detected in all of the core-truncated constructs tested, but readily detected in constructs encoding full-length core sequences. CONCLUSION: The upstream conserved full-length core coding sequence was required for the production of E2 glycoproteins carrying complex-type N-glycans which reacted strongly with homologous patient serum and therefore possibly represented more mature forms of E2. As complex-type N-glycans indicated modification by Golgi enzymes, the results suggest that the presence of full-length core might be critical for E1/E2 complex to leave ER. Our data may contribute to a better understanding of the processing of HCV structural proteins as well as HCV morphogenesis.

The characterization of transmembrane protein 59-like(TMEM59L)reveals its role in the regulating the level of the GDI protein family

The characterization and functions of transmembrane protein 59-like(TMEM59L),a type I transmembrane protein,are not clearly understood until now.Some TMEM59L and fluorescent fusion proteins constructs were transfected in cell lines and liposomes,and their localization was observed.The effects of protein constructs were studied by fluorescence microscopy and western blotting.This study reports a novel function of human TMEM59L(hTMEM59L)related to the expression and location of some proteins.In addition,we report two novel splice variants of human TMEM59L(hTMEM59L).The localization of mutants of this protein,lacking a middle region,and a Cterminal deletion,markedly differed from that of full-length hTMEM59L.Intracellular movement assessment in living cells showed the localization of TMEM59L to vesicular structures in the Golgi bodies,and the cell membrane was observed in living cells.Overexpression of TMEM59L markedly increased the level of amyloid precursor protein because of TMEM59L-mediated inhibition of cell membrane transport but did not affect the expression of betasecretase 2.TMEM59L overexpression also significantly increased the levels of Rab GDP dissociation inhibitor alpha and Rab GDP dissociation inhibitor beta proteins.These results suggest that TMEM59L is involved in the packaging of acidic vesicles and thereby functions in the trafficking and processing of intracellular proteins.

The Role of Vacuolar Processing Enzyme

The classification, characters and maturation methods of VPEs were sum- marized, and its regulation function to vacuolar was also analyzed. Furthermore, effects of the enzyme in vacuolar-mediated plant defense mechanism were discussed to point out that VPEs were divided into 3 subfamilies via autocatalytic mature including seed-type VPE, vegetative-type VPE and new-type VPE. Especially, seed- type VPE mediated the process of storage protein, while vegetative-type VPE and new-type VPE regulated and controlled programmed death of plant cells.

Effect of the desolventizing/toasting process on chemical composition and protein quality of rapeseed meal

Background： During processing in a desolventizer/toaster（DT）, rapeseed meal（RSM） is heated to evaporate the hexane and to reduce the level of heat-labile anti-nutritional factors such as glucosinolates（GSL）. However, excessive heat treatment may reduce amino acid（AA） content in addition to lower AA digestibility and availability in RSM. The objective of the present study was to produce from one batch of a 00-rapeseed variety（17 μmol GSL/g dry matter（DM）, seed grade quality） five differently processed RSM under standardized and defined conditions in a pilot plant,and to determine the impact of these different treatments on protein solubility and chemical composition, in particular with regard to contents of AA including reactive Lys（rL ys） and levels of total and individual GSL.Methods： Four RSM were exposed to wet toasting conditions（WetT C） with increasing residence time in the DT of 48,64, 76, and 93 min. A blend of these four RSM was further processed, starting with saturated steam processing（〈 100 °C）and followed by exposure to dry toasting conditions（DryT C） to further reduce the GSL content in this RSM.Results： The contents of neutral detergent fiber and neutral detergent fiber bound crude protein（CP） increased linearly（P 〈 0.05）, as residence time of RSM in the DT increased from 48 to 93 min, whereas contents of total and most individual GSL and those of Lys, rL ys, Cys, and the calculated ratio of Lys：CP and r Lys：CP decreased linearly（P ≤ 0.05）.The combination of wet heating and DryT C resulted in the lowest GSL content compared to RSM produced under WetT C, but was associated with lowest protein solubility.Conclusions： It can be concluded that by increasing residence time in the DT or using alternative processing conditions such as wet heating combined with DryT C, contents of total and individual GSL in RSM can be substantially reduced.Further in vivo studies are warranted to elucidate if and to which extent the observed differences in protein quality and GSL content between RSM may affect digestibility and bioavailability of AA in monogastric animals.

Label-Free and Real-Time Monitor of Binding and Dissociation Processes between Protein A and Swine IgG by Oblique-Incidence Reflectivity Difference Method

Life science has a need for detection methods that are label-free and real-time. In this paper, we have selected staphylococcal protein A （SPA） and swine immunoglobulin G （IgG）, and monitor the bindings between SPA and swine IgG with different concentrations, as well as the dissociations of SPA-swine IgG complex in different pH values of phosphate buffer by oblique-incidence reflectivity difference （OIRD） in a label-free and real-time fashion. We obtain the ON and OFF reaction dynamic curves corresponding to the bindings and dissociations of SPA and swine IgG. Through our analysis of the experimental results, we have been able to obtain the damping coefficients and the dissociation time of SPA and swine IgG for different pH values of the phosphate buffer. The results prove that the OIRD technique is a competing method for monitoring the dynamic processes of biomolecule interaction and achieving the quantitative information of reaction kinetics.

Analysis of differentially expressed genes in the sea cucumber Apostichopus japonicus under heat stress

The sea cucumber Apostichopus japonicus plays important roles in marine benthic ecosystem as environmental cleaners,and it is the important aquaculture species in China.High water temperature poses critical threat for the survival of A.japonicus,which has resulted in extensive death in summer.To explore the genes expression profiles under different levels of heat stress,the high-throughput RNA-seq was applied in this study.Our results revealed a total of 1371,1225 and 1408 differentially expressed genes(DEGs)in 26℃for 6 h,26℃for 48 h and 30℃for 6 h respectively in comparison with Control group.The pathway analysis suggested“Protein processing in endoplasmic reticulum(ER)”was significantly enriched in all these heat stress(HS)treatment groups.The expression results of key DEGs in this pathway(Hsp70,Derlin,NEF,PDI,GPR94 and ERP57)by qRT-PCR was in accordance with the RNA-seq data.The subcluster analysis of DEGs revealed that a variety of heat shock proteins(HSPs)and calcium ion binding proteins had an obvious up-regulated expression in 26℃for 6 h,comparatively low expression in 26℃for 48 h,and the highest expression in 30℃for 6 h.The other DEGs subcluster,consisting of critical components of extracellular matrix(ECM)and a subset of peptidases and proteases,showed significantly rising tendency in 30℃for 6 h.Additionally,the expression of matrix metalloproteases(MMP1,MMP16 and MMP19)was prominently affected by HS,and peaked in 30℃for 6 h.This study provides a series of candidate genes for further study about heat shock response in A.japonicus,especially genes associated with protein processing in ER and regulation of ECM,which also offers new insights into cellular homeostasis under stressful conditions in marine invertebrates.

Common therapeutic target for both cancer and obesity

Obesity and cancer are two interrelated conditions of high epidemiological need, with studies showing that obesity is responsible for nearly 25% of the relative contribution to cancer incidence. Given the connection between these conditions, a drug that can operate on both obesity and cancer is highly desirable. Such a drug is accomplishablethrough the development of potent anti-angiogenesis agents due to the shared underlying role of angiogenesis in the development of both diseases. Prior research has demonstrated a key role of type-2 methionine aminopeptidase(Met AP2) for angiogenesis, which has led to the development of numerous of novel inhibitors. Several irreversible Met AP2 inhibitors have entered clinical trials without great success. Though this lack of success could be attributed to off-target adverse effects, the underlying causes remain unclear. More promising reversible inhibitors have been recently developed with excellent pre-clinical results. However, due to insufficient knowledge of the biological functions of N-terminal protein processing, it is hard to predict whether these novel inhibitors would successfully pass clinical trials and thereby benefit cancer and obesity patients. Significantly more efforts are needed to advance our understanding of the regulation of methionine aminopeptidases and the processes by which they govern the function of proteins.

Sorting and Processing of the Alzheimer's Disease Amyloid Precursor Protein Mediated by the AP-4 Complex

Proteolytic processing of the transmembrane amyloid precursor protein (APP) to aggregation-prone amyloid-β (Aβ) peptide underlies the development of Alzheimer’s disease.

Visualization of amino acid composition differences between processed protein from different animal species by self-organizing feature maps

Amino acids are the dominant organic components of processed animal proteins,however there has been limited investigation of differences in their composition between various protein sources.Information on these differences will not only be helpful for their further utilization but also provide fundamental information for developing species-specific identification methods.In this study,self-organizing feature maps(SOFM) were used to visualize amino acid composition of fish meal,and meat and bone meal(MBM) produced from poultry,ruminants and swine.SOFM display the similarities and differences in amino acid composition between protein sources and effectively improve data transparency.Amino acid composition was shown to be useful for distinguishing fish meal from MBM due to their large concentration differences between glycine,lysine and proline.However,the amino acid composition of the three MBMs was quite similar.The SOFM results were consistent with those obtained by analysis of variance and principal component analysis but more straightforward.SOFM was shown to have a robust sample linkage capacity and to be able to act as a powerful means to link different sample for further data mining.