Comprehensive proteomic analysis of mineral nanoparticles derived from human body fluids and analyzed by liquid chromatography-tandem mass spectrometry |
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| Authors: | Martel Jan Young David Young Andrew Wu Cheng-Yeu Chen Chi-De Yu Jau-Song Young John D |
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| Affiliation: | aLaboratory of Nanomaterials, Chang Gung University, Gueishan, Taoyuan 333, Taiwan, ROC;bGraduate Institute of Biomedical Sciences, Chang Gung University, Gueishan, Taoyuan 333, Taiwan, ROC;cCenter for Molecular and Clinical Immunology, Chang Gung University, Gueishan, Taoyuan 333, Taiwan, ROC;dDepartment of Materials Science and Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA;eResearch Center of Bacterial Pathogenesis, Chang Gung University, Gueishan, Taoyuan 333, Taiwan, ROC;fProteomics Core Laboratory, Chang Gung University, Gueishan, Taoyuan 333, Taiwan, ROC;gDepartment of Cell and Molecular Biology, Chang Gung University, Gueishan, Taoyuan 333, Taiwan, ROC;hLaboratory of Cellular Physiology and Immunology, Rockefeller University, New York, NY 10021, USA;iBiochemical Engineering Research Center, Mingchi University of Technology, Taishan, Taipei 24301, Taiwan, ROC |
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| Abstract: | Mineralo-protein nanoparticles (NPs) formed spontaneously in the body have been associated with ectopic calcifications seen in atherosclerosis, chronic degenerative diseases, and kidney stone formation. Synthetic NPs are also known to become coated with proteins when they come in contact with body fluids. Identifying the proteins found in NPs should help unravel how NPs are formed in the body and how NPs in general, be they synthetic or naturally formed, interact within the body. Here, we developed a proteomic approach based on liquid chromatography (LC) and tandem mass spectrometry (MS/MS) to determine the protein composition of carbonate-apatite NPs derived from human body fluids (serum, urine, cerebrospinal fluid, ascites, pleural effusion, and synovial fluid). LC–MS/MS provided not only an efficient and comprehensive determination of the protein constituents, but also a semiquantitative ranking of the identified proteins. Notably, the identified NP proteins mirrored the protein composition of the contacting body fluids, with albumin, fetuin-A, complement C3, α-1-antitrypsin, prothrombin, and apolipoproteins A1 and B-100 being consistently associated with the particles. Since several coagulation factors, calcification inhibitors, complement proteins, immune regulators, protease inhibitors, and lipid/molecule carriers can all become NP constituents, our results suggest that mineralo-protein complexes may interface with distinct biochemical pathways in the body depending on their protein composition. We propose that LC–MS/MS be used to characterize proteins found in both synthetic and natural NPs. |
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| Keywords: | Ectopic calcification LC&ndash MS/MS Mineral nanoparticles Proteomics Protein corona |
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