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Study on screening potential allergenic proteins from infant milk powders based on human mast cell membrane chromatography and histamine release assays

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Journal of Pharmaceutical Analysis 2019年第1期9卷 55-61页

作者： Ping Zhang Yingdi Shi Xiaoshuang He Wei Sun Yanni Lv Xiaofang Hou School of Pharmacy Xi'an Jiaotong University Linyi Institute for Food and Drug Control Shaanxi Engineering Research Center of Cardiovascular Drugs Screening & Analysis

Cow's milk allergy is mainly observed in infants and young children. Most allergic reactions affect the skin, followed by the gastrointestinal and respiratory systems. Conventional diagnosis is based on positive allergy studies and evaluation of parameters including IgE and IgG1 levels, acute allergic skin response and anaphylactic shock reactions. We developed a cell membrane chromatographic(CMC)method based on human mast cells(HMC-1) for screening potential allergens in infant formula milk powders(IFMP). HMC-1 cell membranes were extracted and mixed with silica to prepare cell membrane chromatography columns(10 mm ? 2 mm i.d., 5 mm). Under the conditions of 0.2 mL/min flow rate and214 nm detection wavelength, human breast milk showed no retention. However, IFMP showed clear retention. The retained fractions were collected and analyzed through matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Four major milk proteins, i.e., α-casein, β-casein, α-lactalbumin, and β-lactoglobulin A, were identified. Furthermore, these proteins and β-lactoglobulin B showed clear retention on HMC-1/CMC columns. To test the degranulation effects of the five proteins, histamine and β-hexosaminidase release assays were carried out. All five proteins induced HMC-1 cells to release histamine and β-hexosaminidase. Also, we established a reversed phase liquid chromatographic(RPLC) method for the determination of the five proteins in IFMP and the results showed that 90% proteins in IFMP were α-casein and β-casein. We concluded that cow's milk proteins may be potential allergens and caseins cause more β-casein allergic risk than other proteins. This conclusion was consistent with other studies.

关键词： Allergenic proteins Cell membrane chromatography Milk powders

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Plant peroxisomal solute transporter proteins

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Journal of Integrative Plant Biology 2019年第7期61卷 817-835页

作者： Lennart Charton Anastasija Plett Nicole Linka Institute for Plant Biochemistry and Cluster of Excellence on Plant Sciences (CEPLAS) Heinrich Heine University

Plant peroxisomes are unique subcellular organelles which play an indispensable role in several key metabolic pathways, including fatty acid b-oxidation,photorespiration, and degradation of reactive oxygen species. The compartmentalization of metabolic pathways into peroxisomes is a strategy for organizing the metabolic network and improving pathway efficiency. An important prerequisite, however, is the exchange of metabolites between peroxisomes and other cell compartments. Since the first studies in the 1970s scientists contributed to understanding how solutes enter or leave this *** review gives an overview about our current knowledge of the solute permeability of peroxisomal membranes described in plants, yeast, mammals and other eukaryotes. In general, peroxisomes contain in their bilayer membrane specific transporters for hydrophobic fatty acids(ABC transporter) and large cofactor molecules(carrier for ATP, NAD and CoA). Smaller solutes with molecular masses below 300–400 Da, like the organic acids malate, oxaloacetate, and 2-oxoglutarate, are shuttled via non-selective channels across the peroxisomal *** comparison to yeast, human, mammals and other eukaryotes, the function of these known peroxisomal transporters and channels in plants are discussed in this review.

关键词： Plant peroxisomal solute transporter proteins

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Auto-regulatory feedback by RNA-binding proteins

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Journal of Molecular Cell Biology 2019年第10期11卷 930-939页

作者： Michaela Muller-McNicoll Oliver Rossbach Jingyi Hui Jan Medenbach Institute of Cell Biology and Neuroscience Goethe University FrankfurtMax-von-Laue-Strasse 13D-60438 Frankfurt am MainGermany Institute of Biochemistry Justus-Liebig-University GiessenHeinrich-Buff-Ring 17D-35392 GiessenGermany State Key Laboratory of Molecular Biology CAS Center for Excellence in Molecular Cell ScienceShanghai Institute of Biochemistry and Cell BiologyChinese Academy of SciencesShanghai 200031China Institute of Biochemistry I University of RegensburgUniversitaetsstrasse 31D-93053 RegensburgGermany

RNA-binding proteins(RBPs)are key regulators in post-transcriptional control of gene *** that alter their activity or abundance have been implicated in numerous diseases such as neurodegenerative disorders and various types of *** highlights the importance of RBP proteostasis and the necessity to tightly control the expression levels and activities of *** many cases,RBPs engage in an auto-regulatory feedback by directly binding to and influencing the fate oftheirown mRNAs,exerting control over their own *** this feedback control,RBPs employ a variety of mechanisms operating at all levels of posttranscriptional regulation of gene *** we review RBP-mediated autogenous feedback regulation that either serves to maintain protein abundance within a physiological range(by negative feedback)or generates binary,genetic on/off switches important for *** fate decisions(by positive feedback).

关键词： autogenous regulation protein homeostasis RNA-binding proteins post-transcriptional regulation of gene expressi on

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Oral delivery of proteins and peptides:Challenges, status quo and future perspectives

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Acta Pharmaceutica Sinica B 2021年第8期11卷 2416-2448页

作者： Quangang Zhu Zhongjian Chen Pijush Kumar Paul Yi Lu Wei Wu Jianping Qi Shanghai Skin Disease Hospital Tongji University School of MedicineShanghai 200443China Key Laboratory of Smart Drug Delivery of MOE School of PharmacyFudan UniversityShanghai 201203China Department of Pharmacy Gono Bishwabidyalay(University)Mirzanagar SavarDhaka 1344Bangladesh

proteins and peptides(PPs)have gradually become more attractive therapeutic molecules than small molecular drugs due to their high selectivity and efficacy,but fewer side *** to the poor stability and limited permeability through gastrointestinal(GI)tract and epithelia,the therapeutic PPs are usually administered by parenteral *** the big demand for oral administration in clinical use,a variety of researches focused on developing new technologies to overcome GI barriers of PPs,such as enteric coating,enzyme inhibitors,permeation enhancers,nanoparticles,as well as intestinal *** new technologies have been developed under clinical trials and even on the *** review summarizes the history,the physiological barriers and the overcoming approaches,current clinical and preclinical technologies,and future prospects of oral delivery of PPs.

关键词： proteins Peptides Oral delivery Permeation enhancer Enzyme inhibitor Stability Clinical

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Reduced semen quality in patients with testicular cancer seminoma is associated with alterations in the expression of sperm proteins

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Asian Journal of Andrology 2020年第1期22卷 88-93页

作者： Tania R Dias Ashok Agarwal Peter N Pushparaj Gulfam Ahmad Rakesh Sharma American Center for Reproductive Medicine Cleveland ClinicClevelandOH 44195USA Department of Health Sciences Faculty of Health SciencesUniversity of Beira InteriorCovilha 6201-001Portugal Department of Microscopy and Unit for Multidisciplinary Research in Biomedicine Institute of Biomedical Sciences Abel SalazarUniversity of PortoPorto 4050-313Portugal Center of Excellence in Genomic Medicine Research Faculty of Applied Medical SciencesJeddah 21589Saudi Arabia Division of Pathology School of Medical SciencesSydney UniversityLidcombe NSW 2141Australia

Testicular cancer seminoma is one of the most common types of cancer among men of reproductive *** with this condition usually present reduced semen quality,even before initiating cancer ***,the underlying mechanisms by which testicular cancer seminoma affects male fertility are largely *** aim of this study was to investigate alterations in the sperm proteome of men with seminoma undergoing sperm banking before starting cancer therapy,in comparison to healthy proven fertile men(control group).A routine semen analysis was conducted before cryopreservation of the samples(n=15 per group).Men with seminoma showed a decrease in sperm motility(P=0.019),total motile count(P=0.001),concentration(P=0.003),and total sperm count(P=0.001).Quantitative proteomic analysis identified 393 differentially expressed proteins between the study *** proteins involved in spermatogenesis,sperm function,binding of sperm to the oocyte,and fertilization were selected for validation by western *** confirmed the underexpression of heat shock-related 70 kDa protein 2(P=0.041),ubiquinol-cytochrome C reductase core protein 2(P=0.026),and testis-specific sodium/potassium-transporting ATPase subunit alpha-4(P=0.016),as well as the overexpression of angiotensin I converting enzyme(P=0.005)in the seminoma *** altered expression levels of these proteins are associated with spermatogenesis dysfunction,reduced sperm kinematics and motility,failure in capacitation and *** findings of this study may explain the decrease in the fertilizing ability of men with seminoma before starting cancer therapy.

关键词： male fertility proteomics seminoma sperm proteins sperm quality testicular cancer

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The 2018 Nobel Prize in Chemistry: Engineering proteins(enzymes/peptide/antibodies) towards desired properties via the construction of random libraries

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Science China(Life Sciences) 2019年第5期62卷 713-724页

作者： Zengyi Chang School of Life Sciences State Key Laboratory of Protein and Plant Engineering Research Center for Protein Science Center of History and Philosophy of Science Peking University

The 2018 Nobel Prize in Chemistry was awarded one half to Dr. Frances Arnold (of California Institute of Technology,USA) for her work on 'directed evolution of enzymes', the other half to Dr. George Smith (of the University of Missouri, USA) and Sir Gregory Winter (of MRC Laboratory of Molecular Biology, UK) for their work on 'phage display of peptides and antibodies'.

关键词： The 2018 Nobel Prize in Chemistry towards desired properties via the construction of random libraries Engineering proteins

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Preliminary investigation of sequence-independent DNA binding proteins in rat skeletal muscle sarcoplasmic reticulum and their function

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Science China(Life Sciences) 2000年第2期43卷 209-216页

作者：赵文姜志胜倪菊华陈光慧刘乃奎汤健贾弘褆唐朝枢 1. Institute of Cardiovascular Research Beijing Medical University 100083 Beijing China 2. Institute of Cardiovascular Research The First Hospital of Beijing Medical University 100034 Beijing China 3. Department of Biochemistry and Molecular Biology Beijing Medical University 100083 Beijing China

To observe the binding of plasmid DNA to non-nuclear DNA binding proteins in sar-coplasmic reticulum (SR) and the effects of this binding on SR function, sarcoplasmic reticulum proteins in rat skeletal muscle were isolated by differential centrifuge and sucrose density-gradient centrifuge. The results showed that there are two sequence-independent DNA binding proteins in SR proteins, the molecular weights of which are 83 and 58 ku, respectively. Ca2+ uptake and release of SR were remarkably promoted by the binding of plasmid DNA to DNA binding proteins in SR, the mechanism is probably through increasing of Ca2+-ATPase activity in SR and changing of character of Ca2+ release channel ryanodine receptors induced by the binding. These results suggest that there exist DNA binding proteins in SR and its binding to DNA may affect Ca2+ transport of SR.

关键词： DNA binding proteins of SR Ca2+ uptake and release Ca2+-ATPase ryanodine receptor.

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Proteomics Study of COI1-regulated proteins in Arabidopsis Flower

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Journal of Integrative Plant Biology 2010年第4期52卷 410-419页

作者： Lingling Chua Xiaoyi Shan Junxia Wang Wen Peng Guiyou Zhang Daoxin Xie Institute of Molecular and Cell Biology 61 Biopolis Drive Singapore 138673 protein Science Laboratory of the Ministry of Education School of Life Sciences Tsinghua University Beijing 100084 China

Jasmonates （JAs） are a new class of plant hormone that regulate expression of diverse genes to mediate various plant responses. The Arabidopsis F-box protein COI1 is required for plant defense and male fertility in JA signal pathway. To further investigate the regulatory role of COI1 in male fertility, we compared the proteomics profiles of Arabidopsis wild type （WT） flowers with coi1-1 mutant male-sterile flowers using two-dimensional difference gel electrophoresis coupled with matrix-assisted laser desoption/ionization-time-of-flight mass spectrometry. Sixteen proteins with potential function in specific biological processes such as metabolism processes and defense/stress responses were differentially expressed in WT and coi1-1 mutant flowers. Verification on a phi class glutathione transferase AtGSTF9, one out of these 16 identified proteins, revealed that the expression of AtGSTF9 was severely downregulated in flowers of coi1-1 mutant compared with that of WT. Further function analyses of these genes would provide new insights into the molecular basis of COil-regulated male fertility.

关键词： PROTEOMICS PLANT hormones proteins ARABIDOPSIS INFLORESCENCES

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Selective separation and enrichment of proteins in aqueous two-phase extraction system

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Chinese Chemical Letters 2009年第9期20卷 1100-1102页

作者： Feng Qu Hao Qin Min Dong Dong Xu Zhao Xin Ying Zhao Jing Hua Zhang School of Life Science Beijing Institute of Technology Beijing 100081 China Beijing Center for Physical and Chemical Analysis Beijing 100089 China

A simple aqueous two-phase extraction system （ATPS） of PEG/phosphate was proposed for selective separation and enrichment of proteins. The combination of ATPE with HPLC was applied to identify the partition of proteins in two phases. Five proteins （bovine serum albumin, Cytochrome C, lysozyme, myoglobin, and trypsin） were used as model proteins to study the effect of phosphate concentration and pH on proteins partition. The PEG/phosphate system was firstly applied to real human saliva and plasma samples, some proteins showed obviously different partition in two phases. The primary results manifest the selective separation and enrichment of proteins in ATPS provided the potential for high abundance proteins depletion in proteomics. ~ 2009 Feng Qu. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

关键词： Aqueous two-phase extraction Saliva and plasma proteins Selective separation and enrichment

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Clean SEA-TROSY Experiments to Map Solvent Exposed Amides in Large proteins

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Chinese Journal of Chemistry 2004年第12期22卷 1395-1398页

作者：林东海 Shanghai Institute of Meteria Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 201203 China

It is well known that the SEA-TROSY experiment could alleviate some of the problems of resonance overlap in 15N/2H labeled proteins as it was designed to selectively map solvent exposed amide protons. However, SEA- TROSY spectra may be contaminated with exchange-relayed NOE contributions from fast exchanged hydroxyl or amine protons and contributions from longitudinal relaxation. Also, perdeuteration of the protein sample is a pre-requisite for this experiment. In this communication, a modified version, clean SEA-TROSY, was proposed to eliminate these artifacts and to allow the experiment to be applied to protonated or partially deuterated proteins and protein complexes.

关键词： proteins solvent accessibility hydrogen exchange TROSY NMR

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