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ナカムラ ノブヒロ
NAKAMURA NOBUHIRO
中村 暢宏 所属 京都産業大学 生命科学部 先端生命科学科 職種 教授 |
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| 言語種別 | 英語 |
| 発行・発表の年月 | 1992 |
| 形態種別 | 研究論文 |
| 査読 | 査読あり |
| 標題 | Purification and characterization of a vimentin-specific protease in mouse myeloid leukemia cells. Regulation during differentiation and identity with cathepsin G |
| 執筆形態 | その他 |
| 掲載誌名 | European Journal of Biochemistry |
| 出版社・発行元 | SPRINGER VERLAG |
| 巻・号・頁 | 205(3),pp.947-954 |
| 著者・共著者 | Nakamura, N.,Tsuru, A.,Hirayoshi, K.,Nagata, K. |
| 概要 | Strong vimentin-degrading activity was found in a mouse myelomonocytic leukemic cell line, Ml. When M1 cells were induced to differentiate into macrophage-like cells, this degrading activity decreased, while expression of the vimentin gene increased as reported previously [Tsuru, A., Nakamura, N., Takayama, E., Suzuki, Y., Hirayoshi, K. and Nagata, K. (1990) J. Cell Biol. 110, 1655 - 1664]. This activity was not due to calpain, which was reported to degrade vimentin, because it was independent of the presence or absence of Ca2+. This activity was revealed to be strongly associated with membranes by differential-centrifugation experiments. To identify this protease, purification of the degradation enzyme was performed. A membrane fraction was prepared and extracted with a buffer containing Triton X-100, then subjected to column chromatography using carboxymethyl-Sepharose and heparin-Sepharose. Quantitative analysis using the purified protease revealed that the specificity of this protease was more than 1000-fold higher for vimentin than for bovine serum albumin, ovalbumin and actin. Four protein bands expressing the activity were finally identified by SDS/PAGE. Amino-terminal sequences of these four proteins were identical, suggesting lower-molecular-mass proteins were degradative products. Furthermore, it was revealed that the sequence had the highest similarity with that of human cathepsin G. This result was consistent with the cathepsin-G-like properties of the purified protease, such as the optimum pH and the specificities for inhibitors. The purified protease degraded a synthetic substrate for cathepsin G, succinyl-alanyl-alanyl-prolyl-phenylalanyl-p-nitroanilide, with a comparable specific activity to human cathepsin G and was specifically detected with anti-(human cathepsin G) serum in immunoblot analysis. The purified protease thus belongs to the 'cathepsin G family', and perhaps is a mouse homologue of human cathepsin G. |
| DOI | 10.1111/j.1432-1033.1992.tb16861.x |
| ISSN | 0014-2956 |
| Put Code(ORCID) | 19809429 |