关节软骨细胞中疏水膜蛋白补体的无标记蛋白质组学分析：一种鉴定膜生物标志物的技术
Label-free proteomic analysis of the hydrophobic membrane protein complement in articular chondrocytes: a technique for identification of membrane biomarkers.
作者信息
Matta Csaba, Zhang Xiaofei, Liddell Susan, Smith Julia R, Mobasheri Ali
机构信息
a Department of Veterinary Preclinical Sciences , School of Veterinary Medicine, Faculty of Health and Medical Sciences, University of Surrey , Guildford , Surrey , UK .
b Department of Anatomy , Histology and Embryology, Faculty of Medicine, University of Debrecen , Debrecen , Hungary .
出版信息
Biomarkers. 2015;20(8):572-89. doi: 10.3109/1354750X.2015.1130191. Epub 2016 Feb 10.
CONTEXT
There is insufficient knowledge about the chondrocyte membranome and its molecular composition.
OBJECTIVE
To develop a Triton X-114 based separation technique using nanoLC-MS/MS combined with shotgun proteomics to identify chondrocyte membrane proteins.
MATERIALS AND METHODS
Articular chondrocytes from equine metacarpophalangeal joints were separated into hydrophobic and hydrophilic fractions; trypsin-digested proteins were analysed by nanoLC-MS/MS.
RESULTS
A total of 315 proteins were identified. The phase extraction method yielded a high proportion of membrane proteins (56%) including CD276, S100-A6 and three VDAC isoforms.
DISCUSSION
Defining the chondrocyte membranome is likely to reveal new biomarker targets for conventional and biological drug discovery.
背景
关于软骨细胞膜组及其分子组成的知识尚不充分。
目的
开发一种基于Triton X-114的分离技术，结合纳升液相色谱-串联质谱法（nanoLC-MS/MS）和鸟枪法蛋白质组学来鉴定软骨细胞膜蛋白。
材料与方法
将马掌指关节的关节软骨细胞分离为疏水和亲水部分；经胰蛋白酶消化的蛋白质通过nanoLC-MS/MS进行分析。
结果
共鉴定出315种蛋白质。相萃取法产生了高比例的膜蛋白（56%），包括CD276、S100-A6和三种电压依赖性阴离子通道（VDAC）异构体。
讨论
定义软骨细胞膜组可能会揭示用于传统和生物药物研发的新生物标志物靶点。
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