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Abstract
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.
Declaration of interest
The authors do not have any commercial relationships that could be construed as biased or inappropriate. The authors report no declarations of interests. C. M. is supported by the European Commission through a Marie Skłodowska-Curie Intra-European Fellowship for career development (project number: 625746; acronym: CHONDRION; FP7-PEOPLE-2013-IEF). A. M. is the co-ordinator of the D-BOARD Consortium funded by European Commission Framework 7 programme (EU FP7; HEALTH.2012.2.4.5-2, project number 305815, Novel Diagnostics and Biomarkers for Early Identification of Chronic Inflammatory Joint Diseases). A. M. has received funding from the Deanship of Scientific Research (DSR), King AbdulAziz University (grant no. 1-141/1434 HiCi). A. M. is also a member of the Arthritis Research UK Centre for Sport, Exercise, and Osteoarthritis, funded by Arthritis Research UK (Grant Reference Number: 20194).