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Label-free quantification proteomics reveals novel calcium binding proteins in matrix vesicles isolated from mineralizing Saos-2 cells

Label-free quantification proteomics reveals novel calcium b...

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第八届全国医学生物化学与分子生物学第五届全国临床应用生物化学与分子生物学2013华东六省一市生物化学与分子生物学联合学术研讨会

作者： Xiaoying Zhou Yazhou Cui Jing Luan Xiaoyan Zhou Genglin Zhang Xiumei Zhang Jinxiang Han Shandong Academy of Medical Sciences Shandong Medical Biotechnological CenterKey Laboratory for Biotech Drugs of the Ministry of Health School of Medicine and Life Sciences University of Jinan-Shandong Academy of Medical Science

matrix vesicles(MVs) involved in the initiation of mineralization by deposition of hydroxyapatite(HA) in their lumen are released by the budding of mineralization-competent cells during skeletogenesis and bone *** identify additional mineralization-related proteins,MVs were isolated from non-stimulated and stimulated Saos-2 cells in culture via an ExoquickTM approach and the corresponding proteomes were identified and quantified with label-free quantitative proteome *** isolated MVs were confirmed by electron microscopy,alkaline phosphatase activity(ALP),biomarkers,and mineral formation ***-free quantitative proteome analysis revealed that 19 calcium binding proteins(CaBPs),including Grp94,calnexin,calreticulin,calmodulin,and S100 A4/A10,were up-regulated in MVs of Saos-2 cells upon stimulation of *** result provides new clues to study the mechanism of the initiation of MV-mediated mineralization.

关键词： Mineralization matrix vesicles ExoquickTM label-free LC-MS/MS calcium-binding Proteins

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Tenascin C Affects Mineralization Via matrix vesicles And The Mechanism In Saos-2 Cells

Tenascin C Affects Mineralization Via Matrix Vesicles And Th...

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第五届泛环渤海生物化学与分子生物学会学术交流会

作者： LI Cheng-Zhi CUI Ya-Zhou LI Hai-Ying WANG Hua-Xin LUAN Jing ZHOU Xiao-Yan HAN Jin-Xiang Key Laboratory for Rare Disease Research of Shandong Province Key Laboratory for Biotech Drugs of the Ministry of HealthShandong Medical Biotechnological CenterShandong Academy of Medical Sciences School of Medicine and Life Sciences University of Jinan-Shandong Academy of Medical Science Shandong University of Traditional Chinese Medicine

Our previous experiment found that tenascin C was increased with the time-dependent mineralization and we found it was expressed in MVs. In this study, we investigated the effect of TNC on Saos-2 cells differentiation, and explored its possible molecular mechanisms. Two si RNA were added in induced Saos-2 cells, cell differentiation was evaluated by ALP activity, mineralized nodules formation, calcium deposition, and expressions of the osteogenic marker genes. Our results indicated that downregulated TNC expression inhibits Saos-2 cells differentiation as evidenced by decreases in ALP activity, mineralized nodule formation, calcium deposition, and down-regulation of the osteogenic marker genes ALP and COL1 A1. Mechanistically, the activity and ability of MVs were decreased by examining the activity of ALP, calcium deposition and annexins located in MVs. Our study suggests that down-regulated TNC expression might suppress the mineralization, and addition of that may have an effect on the MVs. The results may indicate that TNC could regulate mineralization by regulating the activity and ability of MVs.

关键词： matrix vesicles tenascin mineralization

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The roles of matrix vesicles in bone formation and heterotopic ossification

The roles of matrix vesicles in bone formation and heterotop...

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第八届全国医学生物化学与分子生物学第五届全国临床应用生物化学与分子生物学2013华东六省一市生物化学与分子生物学联合学术研讨会

作者： Pinglan Ge Yazhou Cui Jing Luan Xiaoyan Zhou Jinxiang Han Key Laboratory for Rare Disease Research of Shandong Province

In normal and healthy individuals,bone formation is normally limited to the specialized tissues,the skeleton and the dentition which forms through endochondral ossification or intramembranous *** these hard tissues,bone formation is tightly controlled both in growth and development and in normal adult *** contrast,heterotopic ossification is a pathological condition in which osteogenesis occurs outside of the skeleton,within soft tissues of the ***,the resulting extraskeletal bone is qualitatively normal endochondral or intramembranous bone,and develops through processes that parallel the events that occur during normal embryonic bone and skeletal formation,and also in bone regeneration during fracture *** increasing evidence supports the contention that the mechanisms of soft tissue ossification are similar to those seen in normal bone *** particular,matrix vesicles(MVs),small(20-200 nm) membrane particles,probably play a prominent role in mineral deposition and skeletal *** small particles are budded off from the plasma membrane of chondrocytes,osteoblasts,and odontoblasts prior to the onset of matrix mineralization and are released into the pre-mineralized organic *** perform specialized roles in initiating matrix mineralization,which regulate the Pi/PPi ratio in the intracellular and extracellular fluid,manage mineral nucleation for hydroxylapatite(HA),control calcium and Pi ion homeostasis,and interact with the surrounding ECM to direct HA localization and *** mechanisms involved in the ossification of mineralized tissues have been under investigation for many years and much progress has been made in this *** how matrix vesicles function in normal ossification and the evidence for their participation in pathologic ossification remains to be further researched and *** purpose of this study is to explore the mechanisms of MVs in hard tissue mineralization and then t

关键词： bone formation heterotopic ossification matrix vesicles mineralization hydroxylapatite

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