Objective: To identify antigenic proteins from the salivary glands of female Anopheles maculatus using a proteomic approach to find the biomarker candidate for serological tools.Methods: The identification of antigeni...Objective: To identify antigenic proteins from the salivary glands of female Anopheles maculatus using a proteomic approach to find the biomarker candidate for serological tools.Methods: The identification of antigenic proteins of Anopheles maculatus salivary gland used these techniques: one-dimensional gel electrophoresis(sodium dodecyl sulfate polyacrylamide gel electrophoresis), western blot, and liquid chromatography–mass spectrometry.Results: The proteins that have molecular weight(MW) 43 and 34 k Da were the antigenic protein. Computational bioinformatic analysis by Mascot Server revealed seven novel hypothetical proteins(MW: 43 k Da) and two novel hypothetical proteins(MW:34 k Da). Further analysis(BLASTP, antigenicity, epitope mapping, and specificity analysis) showed that two novel proteins were identified as apolipoprotein D and cathepsin D in Anopheles darlingi.Conclusions: The identified proteins are potential to be developed as a biomarker of mosquito bite's exposure.展开更多
食品乳液在口腔加工过程中的脂肪感知属性主要来自于其乳液液滴与口腔软表面及唾液之间的复杂相互作用,乳滴在舌头表面的黏附、润湿、铺展等一系列界面行为影响乳液在口腔中的摩擦特性和润滑作用。采用高压微射流处理后的豌豆分离蛋白(p...食品乳液在口腔加工过程中的脂肪感知属性主要来自于其乳液液滴与口腔软表面及唾液之间的复杂相互作用,乳滴在舌头表面的黏附、润湿、铺展等一系列界面行为影响乳液在口腔中的摩擦特性和润滑作用。采用高压微射流处理后的豌豆分离蛋白(pea protein isolate,PPI)作为乳化剂制备具有不同油滴粒径的水包油(O/W)乳液,研究油滴粒径变化及施加法向载荷、唾液添加等对PPI稳定的O/W乳液摩擦学特性的影响。结果表明:随着施加法向荷载的增大(0.5~2.0 N),乳液的摩擦系数明显下降,同时,随着乳液粒径的减小,更多乳滴会夹带到接触面之间,从而导致乳液摩擦系数下降(10%~30%),边界层区范围减小15%~20%,能在较低的滑动速率下实现润滑;在模拟口腔加工过程中,唾液的添加会使乳滴发生絮凝,粒径增大,不易夹带到接触面之间,进而导致PPI乳液摩擦系数增大。本研究可为调控豌豆蛋白乳液的口腔质构感官特性及产品研发提供一定技术支持。展开更多
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 prote...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.展开更多
Age-related NADH oxidase (arNOX = ENOX3) proteins are superoxide-generating cell surface oxidases that increase in activity with age beginning at about 30 y. A soluble and truncated exfoliated form of the activity is ...Age-related NADH oxidase (arNOX = ENOX3) proteins are superoxide-generating cell surface oxidases that increase in activity with age beginning at about 30 y. A soluble and truncated exfoliated form of the activity is present in blood and other body fluids. The activity was purified to apparent homogeneity from human urine and resolved by 2-D gel electrophoresis into a series of 24 to 32 kDa components of low isoelectric point. The purified proteins were resistant both to N-terminal sequencing and trypsin cleavage. Cleavage with pepsin revealed peptides corresponding to the TM9 family of transmembrane proteins. Peptide antisera raised to all five members of the human TM9 family sequentially blocked the arNOX activity of human saliva and sera. The soluble truncated N-terminus of the human homolog TM9SF4 was expressed in bacteria. The recombinant protein was characterized biochemically and exhibited ar-NOX activity. The findings identify five arNOX isoforms each of which correspond to one of the five known TM9 family members. The exfoliated soluble arNOX forms are derived from the 24 to 32 kDa N-termini exposed to the cell’s exterior at the cell surface. Each of the shed forms contain putative functional motifs characteristic of ECTO-NOX (ENOX) proteins despite only minimal sequence identity. Our findings identify arNOX as having functional characteristics of ENOX proteins and the TM9 superfamily of proteins as the genetic origins of the five known arNOX isoforms present in human sera, plasma and other body fluids1.展开更多
基金Funded by a Doctorate Research Grant of the Directorate General of Higher Education Indonesia(Grant No.435/UN25.3.1/LT.6/2014)
文摘Objective: To identify antigenic proteins from the salivary glands of female Anopheles maculatus using a proteomic approach to find the biomarker candidate for serological tools.Methods: The identification of antigenic proteins of Anopheles maculatus salivary gland used these techniques: one-dimensional gel electrophoresis(sodium dodecyl sulfate polyacrylamide gel electrophoresis), western blot, and liquid chromatography–mass spectrometry.Results: The proteins that have molecular weight(MW) 43 and 34 k Da were the antigenic protein. Computational bioinformatic analysis by Mascot Server revealed seven novel hypothetical proteins(MW: 43 k Da) and two novel hypothetical proteins(MW:34 k Da). Further analysis(BLASTP, antigenicity, epitope mapping, and specificity analysis) showed that two novel proteins were identified as apolipoprotein D and cathepsin D in Anopheles darlingi.Conclusions: The identified proteins are potential to be developed as a biomarker of mosquito bite's exposure.
文摘食品乳液在口腔加工过程中的脂肪感知属性主要来自于其乳液液滴与口腔软表面及唾液之间的复杂相互作用,乳滴在舌头表面的黏附、润湿、铺展等一系列界面行为影响乳液在口腔中的摩擦特性和润滑作用。采用高压微射流处理后的豌豆分离蛋白(pea protein isolate,PPI)作为乳化剂制备具有不同油滴粒径的水包油(O/W)乳液,研究油滴粒径变化及施加法向载荷、唾液添加等对PPI稳定的O/W乳液摩擦学特性的影响。结果表明:随着施加法向荷载的增大(0.5~2.0 N),乳液的摩擦系数明显下降,同时,随着乳液粒径的减小,更多乳滴会夹带到接触面之间,从而导致乳液摩擦系数下降(10%~30%),边界层区范围减小15%~20%,能在较低的滑动速率下实现润滑;在模拟口腔加工过程中,唾液的添加会使乳滴发生絮凝,粒径增大,不易夹带到接触面之间,进而导致PPI乳液摩擦系数增大。本研究可为调控豌豆蛋白乳液的口腔质构感官特性及产品研发提供一定技术支持。
基金supported by the National Basic Research Program of China(973 program No.2007CB914101)National Natural Science Foundation of China(No.20875009)
文摘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.
文摘Age-related NADH oxidase (arNOX = ENOX3) proteins are superoxide-generating cell surface oxidases that increase in activity with age beginning at about 30 y. A soluble and truncated exfoliated form of the activity is present in blood and other body fluids. The activity was purified to apparent homogeneity from human urine and resolved by 2-D gel electrophoresis into a series of 24 to 32 kDa components of low isoelectric point. The purified proteins were resistant both to N-terminal sequencing and trypsin cleavage. Cleavage with pepsin revealed peptides corresponding to the TM9 family of transmembrane proteins. Peptide antisera raised to all five members of the human TM9 family sequentially blocked the arNOX activity of human saliva and sera. The soluble truncated N-terminus of the human homolog TM9SF4 was expressed in bacteria. The recombinant protein was characterized biochemically and exhibited ar-NOX activity. The findings identify five arNOX isoforms each of which correspond to one of the five known TM9 family members. The exfoliated soluble arNOX forms are derived from the 24 to 32 kDa N-termini exposed to the cell’s exterior at the cell surface. Each of the shed forms contain putative functional motifs characteristic of ECTO-NOX (ENOX) proteins despite only minimal sequence identity. Our findings identify arNOX as having functional characteristics of ENOX proteins and the TM9 superfamily of proteins as the genetic origins of the five known arNOX isoforms present in human sera, plasma and other body fluids1.
