A Surface Loop in the N-Terminal Domain of Pedobacter heparinus Heparin Lyase Ⅱ is Important for Activity
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- Mori Marina
- Department of Applied Biological Science, Tokyo University of Agriculture and Technology
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- Ichikawa Megumi
- Department of Applied Biological Science, Tokyo University of Agriculture and Technology
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- Kiguchi Yumiko
- Department of Applied Biological Science, Tokyo University of Agriculture and Technology
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- Miyazaki Takatsugu
- Department of Applied Biological Science, Tokyo University of Agriculture and Technology
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- Hattori Makoto
- Department of Applied Biological Science, Tokyo University of Agriculture and Technology
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- Nishikawa Atsushi
- Department of Applied Biological Science, Tokyo University of Agriculture and Technology
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- Tonozuka Takashi
- Department of Applied Biological Science, Tokyo University of Agriculture and Technology
書誌事項
- タイトル別名
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- A Surface Loop in the N-Terminal Domain of <i>Pedobacter heparinus </i>Heparin Lyase II is Important for Activity
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抄録
Pedobacter heparinus heparin lyase II (PhHepII) is composed of N-terminal, central, and C-terminal domains. A long surface loop, designated loop-A, is in the N-terminal domain and is composed of amino acids 84-89. In this study, we deleted two, three, or four residues in loop-A to create Δ86-87, Δ85-87, and Δ84-87 PhHepII deletion mutants. We hypothesized that the deletions would increase PhHepII thermostability. After heating purified PhHepII enzymes at 45 °C for 5 min, 6.1 % of the enzyme activity remained in wild-type PhHepII, whereas 10.6 % of the enzyme activity remained in Δ86-87 PhHepII. The results indicated that the deletion caused a significant decrease in the activity, although Δ86-87 PhHepII is slightly more thermostable than wild-type PhHepII. In addtion, Δ85-87 and Δ84-87 PhHepII had weak or no enzyme activity, even when unheated. Circular dichroism spectra showed that Δ85-87 PhHepII was properly folded. These results suggest that the flexibility of loop-A is important for PhHepII enzyme activity.
収録刊行物
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- Journal of Applied Glycoscience
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Journal of Applied Glycoscience 63 (1), 7-11, 2016
日本応用糖質科学会
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詳細情報 詳細情報について
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- CRID
- 1390282681269984384
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- NII論文ID
- 130005126301
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- NII書誌ID
- AA11809133
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- ISSN
- 18807291
- 13447882
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- NDL書誌ID
- 027145200
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
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