Tandem Mass Spectrometric Characterization of Fetuin Sialylated Glycopeptides Enriched by Ti02 Microcolumn

Sialylation of glycoproteins is vital for the function or physicochemical properties of a protein. It becomes more and more important to develop approaches that can be used to efficiently isolate and identify sialylated glycopep- tides or glycoproteins for monitoring changes in glycoproteome. In the present study, we analyze intact structures of the enriched sialylated glycopeptides of bovine fetuin by matrix-assisted laser desorption/ionization-tandem mass spectrometry （MALDI-MS/MS）, without any chemical derivation. The experimental data show that the optimal loading buffer for TiO2 as matrix is 80% acetonitrile/2% TFA （trifluoroacetic acid）/100 mg/mL DHB （2,5-dihydroxybenzoic acid） which is also compatible with MALDI-mass spectrometric analysis. This study indi- cates that the improved enrichment approach combined with MALDI-MS/MS may be a powerful tool to analyze intact structures and components of the sialylated glycopeptides from complex peptide mixture.

Poplar catkin:A natural biomaterial for highly specific and efficient enrichment of sialoglycopeptides

Sialic acids as terminal entities of larger glycans linked to proteins and lipids are involved in multiple different pathological and physiological processes.Structural characterisation of sialoglycoconjugates is required to understand their biological function.However,a comprehensive sialylation analysis of sialoglycoconjugates has remained challenges.In this study,we employ a natural biomaterial,poplar catkin derived from white poplar tree（Populus tomentosa Carr.）,to develop a novel capturing microtip for selective and efficient enrichment of sialoglycopeptides,without losses of sialic acid residues and water molecules from sialoglycopeptides.Scanning electron microscopy and Fourier-transform infrared spectroscopy analysis,along with Maule and Wiesner staining assays,indicated that the main components on the outer layer of the poplar catkin are syringyl and guaiacyl lignins which play a key role in enriching sialoglycopeptides from complex peptide mixture.

Herbal decoctosome is a novel form of medicine

Traditionally, herbal medicine is consumed by drinking decoctions produced by boiling herbs with water. The functional components of the decoction are heat stable. Small RNAs(sRNAs) were reported as a new class of functional components in decoctions. However, the mechanisms by which sRNAs survive heat treatment of the decoction and enter cells are unclear.Previous studies showed that plant-derived exosome-like nanoparticles(ELNs), which we call botanosomes, could deliver therapeutic reagents in vivo. Here, we report that heat-stable decoctosomes(ELNs) from decoctions have more therapeutic effects than the decoctions in vitro and demonstrate therapeutic efficacy in vivo. Furthermore, sRNAs, such as HJT-sRNA-m7 and PGY-sRNA-6, in the decoctosome exhibit potent anti-fibrosis and anti-inflammatory effects, respectively. Decoctosome is comprised of lipids, chemical compounds, proteins, and s RNAs. A medical decoctosome mimic is called bencaosome. A single lipid sphinganine(d22:0) identified in the decoctosome was mixed and heated with the synthesized sRNAs to form the simplest bencaosome. This simple bencaosome structure was identified by critical micelle concentration(cmc) assay that sRNAs coassembled with sphinganine(d22:0) to form the lipid layers of vesicles. The heating process facilitates co-assembly of sRNAs and sphinganine(d22:0) until a steady state is reached. The artificially produced sphinganine-HJT-sRNA-m7 and sphinganinePGY-sRNA-6 bencaosomes could ameliorate bleomycin-induced lung fibrosis and poly(I:C)-induced lung inflammation, respectively, following oral administration in mice. Our study not only demonstrates that the herbal decoctosome may represent a combinatory remedy in precision medicine but also provides an effective oral delivery route for nucleic acid therapy.

Erratum to:Herbal decoctosome is a novel form of medicine

Following the published article,we noticed an error duplication in Figure 5G“control”and“PGY-6”that was introduced during the revised process,with an attempt to replace it with higher-resolution images.Here we provide the original data in the first submitted manuscript(Figure 5G).

Proteomics-based Characterization of Protein Complexes from Human Pancreatic Cancer Cell Line

为了开发一条节俭、可行的途径探查蛋白质建筑群，微分 centrifugation 和三维的 polyacrylamide，胶化电气泳动(页) 被执行把蛋白质建筑群与人的胰腺的癌症房间线的房间 lysate 分开， SW1990，由集体度谱的鉴定列在后面。四 macromolecular 蛋白质建筑群明白地被分开并且识别。

Encapsidating artificial human papillomavirus-16 mE7 protein in human papillomavirus-6b L1/L2 virus like particles

Background Human papillomaviruses （HPVs） can infect squamous or mucosal epithelia and cause cervical cancer or genital warts. Coinfection with multiple HPV types is a common finding of many epidemiological studies. Therefore, it is necessary to develop a vaccine, which can eradicate established HPV infections and prevent other HPV infections. In this study, we generated chimeric virus like particles （cVLPs） composed of HPV-6b L1, HPV-6b L2 and one artificial HPV-16 mE7 proteins. Methods The artificial HPV-16 mE7 gene was designed by codon modification, point mutation and gene shuffling then chemically synthesized and subcloned behind HPV-6b L2. HPV-6b L1 and L2-mE7 were expressed in insect cells by using Bac-to-Bac system. The generated cVLPs were purified by CsCI gradient ultracentrifuge and analyzed by immunoblot, electron microscope and haemagglutination assay. Results The HPV-6b L1 and L2-mE7 proteins were well expressed in insect cells and could selfassemble into cVLPs, whose diameter was about 55 nm and similar to that of HPV-6b L1/L2 VLPs. Intact cVLPs could be recognized by H6.M48 neutralizing monoclonal antibody and HPV-6b L2 polyclonal antibody, while the denatured cVLPs, but not the intact cVLPs, were reactive to HPV-16 E7 polyclonal antibody. HPV-6b LI/L2-mE7 cVLPs haemaggiutinated mouse erythrocytes as efficiently as HPV-6b L1/L2 VLPs did. Conclusions The insertion of the 158 amino acid HPV-16 mE7 protein behind L2 did not disrupt the correct assembling of cVLPs. The morphological characteristics and haemagglutinating activity of cVLPs were similar to those of HPV-6b LI/L2 VLPs. The cVLPs retained conformational B cell epitopes of HPV-6 VLPs and HPV-16 mE7 protein had an internal location in the cVLPs. Therefore, large modified E7 protein with higher immunogenicity could be incorporated into cVLPs by fusing to the C-terminus of L2, which would help to improve the therapeutic effects of LI/L2-E7 cVLPs.