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Title: Cloning, expression and purification of the anion exchanger 1 homologue from the basidiomycete Phanerochaete chrysosporium.
Authors: Tokuda, Natsuko
Igarashi, Kiyohiko
Shimamura, Tatsuro  kyouindb  KAKEN_id
Yurugi-Kobayashi, Takami
Shiroishi, Mitsunori
Ito, Keisuke
Sugawara, Taishi
Asada, Hidetsugu  kyouindb  KAKEN_id  orcid https://orcid.org/0000-0001-6255-4728 (unconfirmed)
Murata, Takeshi
Nomura, Norimichi  kyouindb  KAKEN_id
Iwata, So  kyouindb  KAKEN_id
Kobayashi, Takuya  kyouindb  KAKEN_id
Author's alias: 岩田, 想
小林, 拓也
Keywords: Anion exchanger
Pichia pastoris
Cloning
Purification
Phanerochaete chrysosporium
Issue Date: Sep-2011
Publisher: Elsevier Inc.
Journal title: Protein expression and purification
Volume: 79
Issue: 1
Start page: 81
End page: 87
Abstract: 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.
Rights: © 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(Published Version): 10.1016/j.pep.2011.04.006
PubMed ID: 21515379
Appears in Collections:Journal Articles

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