Proteomic analysis of articular cartilage vesicles from normal and osteoarthritic cartilage.
Proteomic analysis of articular cartilage vesicles from normal and osteoarthritic cartilage.
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DOI：
10.1002/art.30120
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发表时间：
2011-02
影响因子：
--
通讯作者：
Wakim, Bassam T.
中科院分区：
文献类型：
--
作者：
Rosenthal, Ann K.;Gohr, Claudia M.;Ninomiya, James;Wakim, Bassam T.
关节软骨囊泡（ACV）是在正常关节软骨中发现的细胞外细胞器。虽然它们最初被定义为在骨关节炎（OA）软骨中产生病理性钙晶体的能力，但它们也可以通过RNA和蛋白质的转移改变正常软骨细胞的表型。我们分析了来自正常和OA人软骨的ACV的蛋白质组。从10个正常和10个OA人膝关节软骨中分离ACV。我们使用胶内胰蛋白酶消化、胰蛋白酶肽的nanospray LC-MS/MS，然后搜索Uniprot数据库的适当子集来鉴定ACV蛋白质组。我们使用Holm-Sidak分析进行多重比较测试，进一步鉴定了一组在正常和OA ACV之间显著不同的蛋白质。在ACV中鉴定了超过1700种蛋白质。大约170个蛋白质满足我们的严格标准，即每个蛋白质存在一个以上的代表性肽，并且错误发现率≤ 5%。这些蛋白质包括细胞外基质成分、磷脂结合蛋白、酶和细胞骨架成分，包括肌动蛋白。虽然仅在正常或OA ACV中观察到很少的蛋白质，但免疫球蛋白和补体成分仅存在于OA ACV中。与正常ACV相比，OA ACV显示基质蛋白聚糖减少，TGFβ诱导的蛋白βig-H3、DEL 1、玻连蛋白和丝氨酸蛋白酶HTRA 1增加（p<0.01）。这些发现支持ACV作为关节软骨生理结构的概念。OA ACV的变化在很大程度上是定量的，反映了基质的改变和炎症的存在，而不是组成的根本变化。
Articular cartilage vesicles (ACVs) are extracellular organelles found in normal articular cartilage. While they were initially defined by their ability to generate pathologic calcium crystals in osteoarthritic (OA) cartilage, they also can alter the phenotype of normal chondrocytes through transfer of RNA and protein. We analyzed the proteome of ACVs from normal and OA human cartilage. ACVs were isolated from 10 normal and 10 OA human knee cartilages. We identified the ACV proteomes using in-gel trypsin digestion, nanospray LC-MS/MS of tryptic peptides followed by searching an appropriate subset of the Uniprot database. We further identified a set of proteins that were significantly different between normal and OA ACVs using Holm-Sidak analysis for multiple comparison testing. Over 1700 proteins were identified in ACVs. Approximately 170 proteins satisfied our stringent criteria of having more than one representative peptide per protein present and a false discovery rate ≤ 5%. These proteins included extracellular matrix components, phospholipid binding proteins, enzymes, and cytoskeletal components, including actin. While few proteins were seen exclusively in normal or OA ACVs, immunoglobulins and complement components were present only in OA ACVs. Compared to normal ACVs, OA ACVs displayed decreases in matrix proteoglycans and increases in TGFβ-induced protein βig-H3, DEL1, vitronectin, and serine protease HTRA1 (p<0.01). These findings lend support to the concept of ACVs as physiologic structures in articular cartilage. Changes in OA ACVs are largely quantitative and reflect altered matrix and the presence of inflammation, rather than fundamental changes in composition.
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DOI：
10.1016/j.bbrc.2009.08.038
发表时间：
2009-10-23
影响因子：
3.1
作者：
Mitton, Elizabeth;Gohr, Claudia M.;McNally, Mark T.;Rosenthal, Ann K.
通讯作者：
Rosenthal, Ann K.
影响因子：
4.4
作者：
Attur, MG;Dave, MN;Amin, AR
通讯作者：
Amin, AR
影响因子：
4
作者：
Johnson, Kristen A.;Rose, David M.;Terkeltaub, Robert A.
通讯作者：
Terkeltaub, Robert A.
影响因子：
4.2
作者：
Derfus, B;Kranendonk, S;Ryan, L
通讯作者：
Ryan, L
影响因子：
--
作者：
Jubeck, Brian;Muth, Emily;Gohr, Claudia M.;Rosenthal, Ann K.
通讯作者：
Rosenthal, Ann K.