Proteomic Analysis of Human Tendon and Ligament : Solubilization and Analysis of Insoluble Extracellular Matrix in Connective Tissues

Connective tissues such as tendon, ligament and cartilage are mostly composed of extracellular matrix (ECM). These tissues are insoluble, mainly due to the highly cross-linked ECM proteins such as collagens. Difficulties obtaining suitable samples for mass spectrometric analysis render the application of modern proteomic technologies difficult. Complete solubilization of them would not only elucidate protein composition of normal tissues but also reveal pathophysiology of pathological tissues. Here we report complete solubilization of human Achilles tendon and yellow ligament, which is achieved by chemical digestion combined with successive protease treatment including elastase. The digestion mixture was subjected to liquid chromatography-mass spectrometry. The low specificity of elastase was overcome by accurate mass analysis achieved using FT-ICR-MS. In addition to the detailed proteome of both tissues, we also quantitatively determine the major protein composition of samples, by measuring peak area of some characteristic peptides detected in tissue samples and in purified proteins. As a result, differences between human Achilles tendon and yellow ligament were elucidated at molecular level.

内容要旨・審査要旨・論文本文の公開：
内容要旨・審査要旨：LID201705191001.pdf
論文本文：k3009_fulltext.pdf
著者の申請により要約(2017-05-19公開)に替えて論文全文を公開(2018-06-14)
本論文は, 著者Nori Satoの学位論文として提出され, 学位審査・授与の対象となっている。

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