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Protein electrophoresis

Jinhua Sun, Yinglin Chen, Feiyue Qin, Xueting Guan, Wei Xu, Liangmei Xu
Interferons have attracted considerable attention due to their vital roles in the host immune response and low induction of antibiotic resistance. In this study, total RNA was extracted from spleen cells of chicken embryos inoculated with Newcastle disease vaccine, and the full-length chicken interferon-γ (ChIFN-γ) gene was amplified by RT-PCR. The full complementary DNA sequence of the ChIFN-γ gene was 495 bp long and was cloned into the prokaryotic expression vector pProEX™HTb . The plasmid was transformed into Escherichia coli DH5α and the expression of ChIFN-γ was induced by isopropyl β-D-1-thiogalactopyranoside...
October 17, 2016: Animal Science Journal, Nihon Chikusan Gakkaihō
Hayato Terayama, Shuichi Hirai, Munekazu Naito, Ning Qu, Chiaki Katagiri, Kenta Nagahori, Shogo Hayashi, Hiraku Sasaki, Shota Moriya, Masaki Hiramoto, Keisuke Miyazawa, Naoyuki Hatayama, Zhong-Lian Li, Kou Sakabe, Masayuki Matsushita, Masahiro Itoh
There are various autoimmunogenic antigens (AIs) in testicular germ cells (TGCs) recognized as foreign by the body's immune system. However, there is little information of TGC-specific AIs being available. The aim of this study is to identify TGC-specific AIs. We have previously established that immunization using viable syngeneic TGC can also induce murine experimental autoimmune orchitis (EAO) without using any adjuvant. This study is to identify TGC-specific AIs by TGC liquid chromatography-tandem mass spectrometry analysis, followed by two-dimensional gel electrophoresis that reacted with serum IgG from EAO mice...
October 18, 2016: Scientific Reports
Hye Yeon Koh, Hyun Park, Jun Hyuck Lee, Se Jong Han, Young Chang Sohn, Sung Gu Lee
Psychrobacter sp. PAMC 21119, isolated from Antarctic permafrost soil, grows and proliferates at subzero temperatures. However, its major mechanism of cold adaptation regulation remains poorly understood. We investigated the transcriptomic and proteomic responses of this species to cold temperatures by comparing profiles at -5°C and 20°C to understand how extreme microorganisms survive under subzero conditions. We found a total of 2,906 transcripts and 584 differentially expressed genes (≥ 2 fold, p <0...
October 17, 2016: Environmental Microbiology
Alessio Soggiu, Cristian Piras, Viviana Greco, Paola Devoto, Andrea Urbani, Luigino Calzetta, Marco Bortolato, Paola Roncada
The enzyme 5α-reductase (5αR) catalyzes the conversion of progesterone and testosterone into neuroactive steroids implicated in a wide array of behavioral functions. The prototypical 5αR inhibitor, finasteride (FIN), is clinically approved for the treatment of androgenic alopecia and benign prostatic hyperplasia. Recent evidence has shown that FIN, albeit generally well tolerated, can induce untoward psychological effects in a subset of patients; furthermore, this drug may have therapeutic efficacy for a number of different neuropsychiatric conditions, ranging from Tourette syndrome to schizophrenia...
October 6, 2016: Psychoneuroendocrinology
Hari Bhupathi Krishnan, Alaa Alaswad, Nathan Wayne Oehrle, Jason Gillman
Legumes form symbiotic association with soil-dwelling bacteria collectively called rhizobia. This association results in the formation of nodules, unique plant-derived organs, within which the rhizobia are housed. Rhizobia encoded-nitrogenase facilitates the conversation of atmospheric nitrogen into ammonia, which is utilized by the plants for its growth and development. Fatty acids have been shown to play an important role in root nodule symbiosis. In this study, we have investigated the role of Stearoyl-Acyl Carrier Protein Desaturase- isoform C (SACPD-C), a soybean enzyme that catalyzes the conversion of stearic acid into oleic acid, which is expressed in developing seeds and in nitrogen fixing nodules...
October 17, 2016: Molecular Plant-microbe Interactions: MPMI
Leslie Muller, Luc Fornecker, Alain Dorsselaer, Sarah Cianférani, Christine Carapito
Sample preparation, typically by in-solution or in-gel approaches, has a strong influence on the accuracy and robustness of quantitative proteomics workflows. The major benefit of in-gel procedures is their compatibility with detergents (such as SDS) for protein solubilization. However, SDS-PAGE is a time-consuming approach. Tube-gel (TG) preparation circumvents this drawback as it involves directly trapping the sample in a polyacrylamide-gel matrix without electrophoresis. We report here the first global label-free quantitative comparison between TG, stacking gel (SG), and basic liquid digestion (LD)...
October 17, 2016: Proteomics
Peizhen Li, Jun Ying, Qingli Chang, Wen Zhu, Guangjian Yang, Teng Xu, Huiguang Yi, Ruowang Pan, Enyong Zhang, Xiaofeng Zeng, Chunxia Yan, Qiyu Bao, Shengbin Li
We studied phycoerythrin (PE) in human SW480 tumor cells and the underlying molecular mechanisms of action. PE inhibited cell proliferation as evidenced by CCK-8 assay. The IC50 values of phycoerythrin were 48.2 and 27.4 µg/ml for 24 and 48 h of exposure, respectively. PE induced apoptosis and cell cycle arrest in SW480 cells as observed under electron microscopy and with flow cytometry. Apoptosis increased from 5.1 (controls) to 39.0% in 80.0 µg/ml PE-treated cells. Differences in protein expression were identified using proteomic techniques...
October 11, 2016: Oncology Reports
Xin Zhou, Ping Wang, Ying-Jian Zhang, Jin-Jin Xu, La-Mei Zhang, Lin Zhu, Li-Ping Xu, Xiao-Min Liu, Huan-Huan Su
The present study aimed to investigate the proteomic difference between melanosis coli (MC) alone and melanosis coli with colon cancer (MCCC). Protein expression in patients with different diseases was analyzed using two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/TOF-MS). A total of 14 protein differences with a confidence level of >95% were found. There were six differences between MC and normal tissues, in which two proteins exhibited upregulated expression levels and four proteins exhibited downregulated expression levels in MC...
October 17, 2016: Oncology Reports
P-H Liao, H-H Hsu, T-S Chen, M-C Chen, C-H Day, C-C Tu, Y-M Lin, F-J Tsai, W-W Kuo, C-Y Huang
Hepatocellular carcinoma (HCC) is the most common type of liver cancer. Despite the availability of several treatment strategies, resistance to chemotherapeutic agents, which limits the effectiveness of anticancer drugs, is a major problem in cancer therapy. In this study, we used a histone deacetylases inhibitor (HDACi) to establish drug-resistant HCC cells and further analyzed the molecular mechanisms underlying the development of resistance in HCC cells. Compared with the parental cells, HDACi-resistant cells showed high metastatic and pro-survival abilities...
October 17, 2016: Oncogene
D S Nunes, H T Gonzaga, V S Ribeiro, J P Cunha-Júnior, J M Costa-Cruz
There is an increasing interest in improving neurocysticercosis (NCC) diagnosis through the search of new and alternative antigenic sources, as those obtained from heterologous antigens. The aim of this study was to obtain potential biomarkers for NCC diagnosis after gel filtration chromatography [gel filtration fraction (GFF)] from the total saline extract (SE) from Taenia saginata metacestodes, followed by protein identification and application in immunodiagnostic. SE and GFF proteic profiles were characterized in gel electrophoresis, and diagnostic performance was verified by testing 160 serum samples through enzyme-linked immunosorbent assay and immunoblotting...
October 17, 2016: Parasitology
Naveen Maddukuri, Qiyang Zhang, Ning Zhang, Maojun Gong
Laser-induced fluorescence (LIF) detection often requires labelling of analytes with fluorophores; and fast fluorescent derivatization is valuable for high-throughput analysis with flow-gated capillary electrophoresis (CE). Here, we report a fast fluorescein-labelling scheme for amino neurotransmitters, which were then rapidly separated and detected in flow-gated CE. This scheme was based on the reaction between primary amines and o-phthalaldehyde (OPA) in the presence of a fluorescent thiol, 2-[(5-fluoresceinyl)aminocarbonyl]ethyl mercaptan (FACE-SH)...
October 17, 2016: Electrophoresis
Wanqing Liu, Quentin M Anstee, Xiaoliang Wang, Samer Gawrieh, Eric R Gamazon, Shaminie Athinarayanan, Yang-Lin Liu, Rebecca Darlay, Heather J Cordell, Ann K Daly, Chris P Day, Naga Chalasani
The increased expression of PNPLA3(148M) leads to hepatosteatosis in mice. This study aims to investigate the genetic control of hepatic PNPLA3 transcription and to explore its impact on NAFLD risk in humans. Through a locus-wide expression quantitative trait loci (eQTL) mapping in two human liver sample sets, a PNPLA3 intronic SNP, rs139051 A>G was identified as a significant eQTL (p = 6.6×10(-8)) influencing PNPLA3 transcription, with the A allele significantly associated with increased PNPLA3 mRNA. An electrophoresis mobility shift assay further demonstrated that the A allele has enhanced affinity to nuclear proteins than the G allele...
October 13, 2016: Aging
Christina Rösch, Fabian Kratz, Thomas Hering, Simone Trautmann, Natalia Umanskaya, Nils Tippkötter, Christine Müller-Renno, Roland Ulber, Matthias Hannig, Christiane Ziegler
The interplay of albumin (BSA) and lysozyme (LYZ) adsorbed simultaneously on titanium was analyzed by gel electrophoresis and BCA assay. It was found that BSA and lysozyme adsorb cooperatively. Additionally, the isoelectric point of the respective protein influences the adsorption. Also, the enzymatic activity of lysozyme and amylase (AMY) in mixtures with BSA was considered with respect to a possible influence of protein-protein interaction on enzyme activity. Indeed, an increase of lysozyme activity in the presence of BSA could be observed...
September 29, 2016: Colloids and Surfaces. B, Biointerfaces
Rosemeyre A Cordeiro, Daniela Santo, Dina Farinha, Arménio Serra, Henrique Faneca, Jorge F J Coelho
Cationic polymer-based vectors have been considered a promising strategy in gene therapy area due to their inherent ability to condense genetic material and successfully transfect cells. However, they usually exhibit high cytotoxicity. In this work, it is proposed the use of a tailor-made gene carrier based on a tri-block copolymer of poly[2-(dimethylamino)ethyl methacrylate] and poly(β-amino ester) (PDMAEMA-b-PβAE-b-PDMAEMA), the influence of the PβAE length being assessed. For this purpose, three different block copolymers were prepared varying the molecular weight of this segment...
October 12, 2016: Acta Biomaterialia
Wei Xiong, Gaozhong Shen, Donald A Bryant
The genome of the model cyanobacterium, Synechococcus sp. PCC 7002, encodes two paralogs of CruA-type lycopene cyclases, SynPCC7002_A2153 and SynPCC7002_A0043, which are denoted cruA and cruP, respectively. Unlike the wild-type strain, a cruA deletion mutant is light-sensitive, grows slowly, and accumulates lycopene, γ-carotene, and 1-OH-lycopene; however, this strain still produces β-carotene and other carotenoids derived from it. Expression of cruA from Synechocystis sp. PCC 6803 (cruA 6803) in Escherichia coli strains that synthesize either lycopene or γ-carotene did not lead to the synthesis of either γ-carotene or β-carotene, respectively...
October 14, 2016: Photosynthesis Research
Chiung-Yueh Hsu, Géraldine Gasc, Anne-Aurélie Raymond, Odile Burlet-Schiltz, Hidenari Takahara, Guy Serre, Marie-Claire Méchin, Michel Simon
Hornerin (HRNR) shares numerous features with filaggrin, a key contributor to the epidermal barrier functions. The two proteins display a related structural organization, are expressed by the granular keratinocytes as a large precursor processed by proteolysis, and are cross-linked to the cornified cell envelopes. Two main steps in the metabolism of filaggrin are its deimination and calpain-1 cleavage. Here, using ion-exchange chromatography and two-dimensional gel electrophoresis of human epidermis extracts, we determined that HRNR is deiminated in vivo...
October 11, 2016: Journal of Investigative Dermatology
Bin Qi, Shan Wang, Qiu Wang, He Zhang, Xue-Yuan Bai, Hui-Nan He, Wei-Jie Sun, Li Liu, Da-Qing Zhao
ETHNOPHARMACOLOGICAL RELEVANCE: Panax quinquefolius L. has been used as a proverbial tonic in oriental countries for hundreds of years. It is used as a traditional medicinal herb to nourish vitality. AIM OF THE STUDY: The purpose of our study was to inquiry the activation effects on murine peritoneal macrophages of a novel protein separated from the roots of Panax quinquefolius L. MATERIALS AND METHODS: In our work, a novel protein of the roots of American ginseng (AGNP) was separated and purified from the roots of Panax quinquefolius L...
October 11, 2016: Journal of Ethnopharmacology
Li Ou, Michael J Przybilla, Chester B Whitley
Mucopolysaccharidosis type I (MPS I) is due to deficiency of α-l-iduronidase (IDUA) and subsequent storage of undegraded glycosaminoglycans (GAG). The severe form of the disease, known as Hurler syndrome, is characterized by mental retardation and neurodegeneration of unknown etiology. To identify potential biomarkers and unveil the neuropathology mechanism of MPS I disease, two-dimensional polyacrylamide gel electrophoresis (PAGE) and nanoliquid chromatography-tandem mass spectrometry (nanoLC-MS/MS) were applied to compare proteome profiling of brains from MPS I and control mice (5-month old)...
October 11, 2016: Molecular Genetics and Metabolism
Jutta Speda, Mikaela A Johansson, Uno Carlsson, Martin Karlsson
Enzyme discovery in individual strains of microorganisms is compromised by the limitations of pure culturing. In principle, metaproteomics allows for fractionation and study of different parts of the protein complement but has hitherto mainly been used to identify intracellular proteins. However, the extracellular environment is also expected to comprise a wealth of information regarding important proteins. An absolute requirement for metaproteomic studies of protein expression, and irrespective of downstream methods for analysis, is that sample preparation methods provide clean, concentrated and representative samples of the protein complement...
October 11, 2016: Analytical Biochemistry
Amma Amponsah, Balunkeswar Nayak
The extraction of soy proteins for soy allergen detections is conventionally achieved with PBS buffer for at least 2 h at room temperature or 4 °C. This method has been reported to be inefficient due to time consumption and inadequate protein extraction resulting in false negative allergen detection and mislabeling of foods containing allergenic proteins. This study investigated the application of microwave (MAE) and ultrasound assisted extraction (UAE) techniques to extract and improve recovery of allergens from various soy matrices...
October 14, 2016: Journal of Food Science
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