Proteome analysis of matrix vesicles isolated from femurs of chicken embryo
Proteome analysis of matrix vesicles isolated from femurs of chicken embryo
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DOI：
10.1002/pmic.200700612
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发表时间：
2008-01-01
期刊：
影响因子：
3.4
通讯作者：
Pikula, Slawomir
中科院分区：
文献类型：
--
作者：
Balcerzak, Marcin;Malinowska, Agata;Pikula, Slawomir
基质小泡(MVS)是启动矿物形成的细胞外细胞器，积累无机磷(P-I)和钙导致羟基磷灰石(HA)晶体的形成，羟基磷灰石(HA)是骨骼的主要矿物成分。MVS在骨形成过程中产生，也在软骨内软骨钙化过程中产生。MVS从成骨细胞和肥大的软骨细胞等骨细胞释放到细胞外基质中。本文采用一维SDS-PAGE、凝胶内胰酶消化和LC-MS-MS/MS蛋白质鉴定方法，对鸡胚骨和软骨中MVS的蛋白质组进行了研究。我们鉴定了126种基因产物，包括与细胞外基质和离子转运相关的蛋白质，以及酶、细胞骨架和调节蛋白。在MVS中首次识别的蛋白质包括水通道蛋白1、膜联蛋白A1(AnxA1)、AnxA11、糖蛋白HT7、G(I)蛋白α2和B型清道夫受体。本文讨论了识别的蛋白质靶向MVS的途径及其在生物矿化过程中的特殊功能。了解这些蛋白质在胚胎矿化过程中的功能和作用是全面了解初始矿物形成机制的先决条件。
Matrix vesicles (MVs) are extracellular organelles that initiate mineral formation, accumulating inorganic phosphate (P-i) and calcium leading to the formation of hydroxyapatite (HA) crystals, the main mineral component of bones. MVs are produced during bone formation, as well as during the endochondral calcification of cartilage. MVs are released into the extracellular matrix from osseous cells such as osteoblasts and hypertrophic chondrocytes. In this report, using 1-D SDS-PAGE, in-gel tryptic digestion and an LC-MS-MS/MS protein identification protocol, we characterized the proteome of MVs isolated from chicken embryo (Gallus gallus) bones and cartilage. We identified 126 gene products, including proteins related to the extracellular matrix and ion transport, as well as enzymes, cytoskeletal, and regulatory proteins. Among the proteins recognized for the first time in MVs were aquaporin 1, annexin A1 (AnxA1), AnxA11, glycoprotein HT7, G(i) protein alpha 2, and scavenger receptor type B. The pathways for targeting the identified proteins into MVs and their particular functions in the biomineralization process are discussed. Obtaining a knowledge of the functions and roles of these proteins during embryonic mineralization is a prerequisite for the overall understanding of the initial mineral formation mechanisms.