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タイトル: | Cloning, expression and purification of the anion exchanger 1 homologue from the basidiomycete Phanerochaete chrysosporium. |
著者: | Tokuda, Natsuko Igarashi, Kiyohiko Shimamura, Tatsuro Yurugi-Kobayashi, Takami Shiroishi, Mitsunori Ito, Keisuke Sugawara, Taishi Asada, Hidetsugu https://orcid.org/0000-0001-6255-4728 (unconfirmed) Murata, Takeshi Nomura, Norimichi https://orcid.org/0000-0002-6330-2239 (unconfirmed) Iwata, So Kobayashi, Takuya |
著者名の別形: | 岩田, 想 小林, 拓也 |
キーワード: | Anion exchanger Pichia pastoris Cloning Purification Phanerochaete chrysosporium |
発行日: | Sep-2011 |
出版者: | Elsevier Inc. |
誌名: | Protein expression and purification |
巻: | 79 |
号: | 1 |
開始ページ: | 81 |
終了ページ: | 87 |
抄録: | Anion exchangers are membrane proteins that have been identified in a wide variety of species, where they transport Cl(-) and HCO3(-) across the cell membrane. In this study, we cloned an anion-exchange protein from the genome of the basidiomycete Phanerochaete chrysosporium (PcAEP). PcAEP is a 618-amino acid protein that is homologous to the human anion exchanger (AE1) with 22.9% identity and 40.3% similarity. PcAEP was overexpressed by introducing the PcAEP gene into the genome of Pichia pastoris. As a result, PcAEP localized in the membrane of P. pastoris and was solubilized successfully by n-dodecyl-β-d-maltoside. His-tagged PcAEP was purified as a single band on SDS-PAGE using immobilized metal affinity chromatography and gel filtration chromatography. Purified PcAEP was found to bind to SITS, an inhibitor of the AE family, suggesting that the purified protein is folded properly. PcAEP expressed and purified using the present system could be useful for biological and structural studies of the anion exchange family of proteins. |
著作権等: | © 2011 Elsevier Inc. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。 This is not the published version. Please cite only the published version. |
URI: | http://hdl.handle.net/2433/143680 |
DOI(出版社版): | 10.1016/j.pep.2011.04.006 |
PubMed ID: | 21515379 |
出現コレクション: | 学術雑誌掲載論文等 |
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