Membrane proteome characterization of periodontal ligament cell sets from deciduous and permanent teeth
Membrane proteome characterization of periodontal ligament cell sets from deciduous and permanent teeth
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DOI：
10.1002/jper.18-0217
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发表时间：
2019-07-01
影响因子：
4.3
通讯作者：
Kantovitz, Kamila R.
中科院分区：
文献类型：
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作者：
Giovani, Priscila A.;Salmon, Cristiane R.;Kantovitz, Kamila R.
牙周韧带（PDL）的生理功能包括牙齿萌出和固支、力吸收和提供本体感觉信息。尽管在了解PDL细胞的生物学方面取得了进展，但缺乏关于落叶（DecPDL）和永久（PermPDL） PDL组织的分子特征的信息。因此，本研究旨在表征DecPDL和PermPDL细胞的膜蛋白质组。方法获得原代PDL细胞（n = 6），对细胞膜富集组分进行无标记定量蛋白质组分析。在新鲜人体组织（n = 8）和原代细胞培养（n = 6）中，通过定量聚合酶链反应和Western blot检测验证了蛋白质组学的发现。此外，用共聚焦显微镜验证目标因子在PDL细胞培养中的表达。结果比较基因本体富集分析表明，大多数粘接差异涉及“膜系统”（PICALM、STX4和LRP10）、“水解酶活性”（NCSTN和XRCC6）、“蛋白结合”（PICALM、STX4、GPNMB、VASP、extend_synaptotagmin 2 [ESYT2]和富含亮氨酸的重复序列15 [LRRC15]）和“异构酶活性”（FKBP8）。数据可通过ProteomeXchange获得，标识符为PXD010226。在转录水平上，在新鲜PDL组织中证实了DecPDL和PermPDL中的ESYT2和LRRC15的高PICALM。此外，Western blot分析证实PICALM、LRRC15和ESYT2在细胞和/或新鲜组织中的表达水平升高，共聚焦显微镜证实了PICALM和LRRC15在PDL细胞中的表达趋势。我们首次全面表征了DecPDL和PermPDL细胞的膜蛋白机制，并共同鉴定了这些细胞群体的独特分子特征，包括DecPDL和PermPDL的独特蛋白质。
Background Physiological roles for the periodontal ligament (PDL) include tooth eruption and anchorage, force absorption, and provision of proprioceptive information. Despite the advances in understanding the biology of PDL cells, there is a lack of information regarding the molecular signature of deciduous (DecPDL) and permanent (PermPDL) PDL tissues. Thus, the present study was designed to characterize the membrane proteome of DecPDL and PermPDL cells. Methods Primary PDL cells were obtained (n = 6) and a label-free quantitative proteome of cell membrane-enriched components was performed. Proteome findings were validated by quantitative polymerase chain reaction and Western blot assays in fresh human tissues (n = 8) and primary cell cultures (n = 6). In addition, confocal microscopy was used to verify the expression of target factors in the PDL cell cultures. Results Comparative gene ontology enrichment analysis evidenced that most stickling differences involved "endomembrane system" (PICALM, STX4, and LRP10), "hydrolase activity" (NCSTN and XRCC6), "protein binding" (PICALM, STX4, GPNMB, VASP, extended-synaptotagmin 2 [ESYT2], and leucine-rich repeat containing 15 [LRRC15]), and "isomerase activity" (FKBP8). Data are available via ProteomeXchange with identifier PXD010226. At the transcript level, high PICALM in DecPDL and ESYT2 and LRRC15 in PermPDL were confirmed in fresh PDL tissues. Furthermore, Western blot analysis confirmed increased levels of PICALM, LRRC15, and ESYT2 in cells and/or fresh tissues, and confocal microscopy confirmed the trends for PICALM and LRRC15 expression in PDL cells. Conclusion We report the first comprehensive characterization of the membrane protein machinery of DecPDL and PermPDL cells, and together, we identified a distinct molecular signature for these cell populations, including unique proteins for DecPDL and PermPDL.