Matsumoto Ken-ichi, Kinoshita Takeshi, Hirose Tomohiro, Ariga Hiroyoshi
Department of Molecular Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
DNA Cell Biol. 2006 Aug;25(8):448-56. doi: 10.1089/dna.2006.25.448.
The interstitial extracellular matrix tenascin-X (iTNX), which has a molecular mass of roughly 450 kDa, is expressed at high levels in muscular tissues and skin. In this study, we identified the serum form of TNX (sTNX) with a molecular mass of 200 kDa in the mouse. Western blot analysis with specific antibodies against fibronectin type III-like (FNIII) repeats of TNX and N-terminal sequence analysis of 200-kDa sTNX revealed that the N-terminus of sTNX is located in the juncture between the 16th FNIII (M16) and 17th FNIII (M17) repeats of iTNX. The 200-kDa sTNX contains 15 FNIII repeats and a fibrinogen domain identical to the Cterminal portion of the iTNX. TNX-deficient mice lacked not only iTNX but also sTNX. Furthermore, 200-kDa sTNX was generated by cleavage of the spleen iTNX by spleen homogenate, and its generation was inhibited by protease inhibitors. These results suggest that sTNX is generated by proteolytic cleavage of iTNX.
分子质量约为450 kDa的间质细胞外基质腱生蛋白X(iTNX)在肌肉组织和皮肤中高水平表达。在本研究中,我们在小鼠体内鉴定出了分子质量为200 kDa的血清形式的TNX(sTNX)。用针对TNX的III型纤连蛋白样(FNIII)重复序列的特异性抗体进行的蛋白质印迹分析以及对200 kDa sTNX的N端序列分析表明,sTNX的N端位于iTNX的第16个FNIII(M16)和第17个FNIII(M17)重复序列之间的连接处。200 kDa的sTNX包含15个FNIII重复序列和一个与iTNX C端部分相同的纤维蛋白原结构域。TNX缺陷小鼠不仅缺乏iTNX,也缺乏sTNX。此外,脾脏匀浆可切割脾脏中的iTNX产生200 kDa的sTNX,蛋白酶抑制剂可抑制其产生。这些结果表明,sTNX是由iTNX的蛋白水解切割产生的。