One-step extraction of functional recombinant aquaporin Z from inclusion bodies with optimal detergent.
Aquaporins are integral membrane channel proteins found in all kingdoms of life. The Escherichia coli aquaporin Z (AqpZ) has been shown to solely conduct water at high permeability. Functional AqpZ is generally purified from the membrane fraction. However, the quantity of the purified protein is limited. In this study, a new method is developed to achieve high yield of bioactive AqpZ protein. A mild detergent n-dodecyl-β-D-maltopyranoside (DDM) was used to solubilize the over-expressed insoluble AqpZ from inclusion bodies without a refolding process. The recovered AqpZ protein showed high water permeability comparable with AqpZ obtained from the membrane fraction. In this way, the total yield of bioactive AqpZ has been increased greatly, which will facilitate the structural and functional characterization and future applications of AqpZ.
NUS Environmental Research Institute (NERI), National University of Singapore, Singapore.
Lili
L
Wang
Department of Biological Sciences, National University of Singapore, Singapore.
Hu
H
Zhou
NUS Environmental Research Institute (NERI), National University of Singapore, Singapore.
Zhengjun
Z
Li
Department of Biological Sciences, National University of Singapore, Singapore.
Teck Kwang
TK
Lim
Department of Biological Sciences, National University of Singapore, Singapore.
Xin Shan
XS
Lim
NUS Environmental Research Institute (NERI), National University of Singapore, Singapore; Department of Biological Sciences, National University of Singapore, Singapore. Electronic address: dbslinqs@nus.edu.sg.
Qingsong
Q
Lin
D020346:Aquaporins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D003902:Detergents / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002479:Inclusion Bodies / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
D020346:Aquaporins; D003902:Detergents; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; C106425:aqpZ protein, E coli
Aquaporin Z; Detergent; Inclusion bodies; Protein purification; Solubility