Immunological detection of alkaline-diaminobenzidine-negative peroxisomes of the nematode Caenorhabditis elegans: Purification and unique pH optima of peroxisomal catalase
European Journal of Biochemistry 267 巻 5 号
1307-1312 頁
2000 発行
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| ファイル情報(添付) | |
| タイトル ( eng ) |
Immunological detection of alkaline-diaminobenzidine-negative peroxisomes of the nematode Caenorhabditis elegans: Purification and unique pH optima of peroxisomal catalase
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| 作成者 |
Togo Summanuna H.
Maebuchi Motohiro
Yokota Sadaki
Bun-ya Masanori
Kawahara Akira
Kamiryo Tatsuyuki
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| 収録物名 |
European Journal of Biochemistry
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| 巻 | 267 |
| 号 | 5 |
| 開始ページ | 1307 |
| 終了ページ | 1312 |
| 抄録 |
We purified catalase-2 of the nematode Caenorhabditis elegans and identified peroxisomes in this organism. The peroxisomes of C. elegans were not detectable by cytochemical staining using 3,3'-diaminobenzidine, a commonly used method depending on the peroxidase activity of peroxisomal catalase at pH 9 in which genuine peroxidases are inactive. The cDNA sequences of C. elegans predict two catalases very similar to each other throughout the molecule, except for the short C-terminal sequence; catalase-2 (500 residues long) carries a peroxisomal targeting signal 1-like sequence (Ser-His-Ile), whereas catalase-1 does not. The catalase purified to near homogeneity from the homogenate of C. elegans cells consisted of a subunit of 57 kDa and was specifically recognized by anti-(catalase-2) serum but not by anti-(catalase-1) serum. Subcellular fractionation and indirect immunoelectron microscopy of the nematode detected catalase-2 inside vesicles judged to be peroxisomes using morphological criteria. The purified enzyme (220 kDa) was tetrameric, similar to many catalases from various sources, but exhibited unique pH optima for catalase (pH 6) and peroxidase (pH 4) activities; the latter value is unusually low and explains why the peroxidase activity was undetectable using the standard alkaline diaminobenzidine- staining method. These results indicate that catalase-2 is peroxisomal and verify that it can be used as a marker enzyme for C. elegans peroxisomes.
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| 著者キーワード |
Caenorhabditis elegans
catalase
3.3'-diaminobenzidine
peroxidase
peroxisomes
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| NDC分類 |
生物科学・一般生物学 [ 460 ]
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| 言語 |
英語
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| 資源タイプ | 学術雑誌論文 |
| 出版者 |
Federation of European Biochemical Societies
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| 発行日 | 2000 |
| 権利情報 |
Copyright (c) 1990 Federation of European Biochemical Societies
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| 出版タイプ | Version of Record（出版社版。早期公開を含む） |
| アクセス権 | オープンアクセス |
| 収録物識別子 |
[ISSN] 0014-2956
[NCID] AA00639541
[PMID] 10691967
[DOI] 10.1046/j.1432-1327.2000.01091.x
[DOI] http://dx.doi.org/10.1046/j.1432-1327.2000.01091.x
～の異版である
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