Crystallization of the membrane protein hVDAC1 produced in cell-free system.
Structural studies of membrane proteins are in constant evolution with the development of new improvements for their expression, purification, stabilization and crystallization. However, none of these methods still provides a universal approach to solve the structure of membrane proteins. Here we describe the crystallization of the human voltage-dependent anion channel-1 produced by a bacterial cell-free expression system. While VDAC structures have been recently solved, we propose an alternative strategy for producing the recombinant protein, which can be applied to other membrane proteins reluctant to expression, purification and crystallization by classical approaches. Despite a lot of efforts to crystallize a cell-free expressed membrane protein, this study is to our knowledge one of the first reports of a successful crystallization. Focusing on expression in a soluble and functional state, in a detergent environment, is the key to get crystals. Although the diffraction of VDAC crystals is limited, the simplicity and the rapidity to set-up and optimize this technology are drastic advantages in comparison to other methods.
Deniaud
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A
CEA, Institut de Biologie Structurale Jean-Pierre Ebel, 41 rue Jules Horowitz 38027 Grenoble, France.
Liguori
L
L
Blesneac
I
I
Lenormand
J L
JL
Pebay-Peyroula
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E
D001483:Base Sequence; D002474:Cell-Free System; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D017931:DNA Primers / Q000235:genetics; D003902:Detergents; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012995:Solubility; D050995:Voltage-Dependent Anion Channel 1 / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D003902:Detergents; D011994:Recombinant Proteins; C494762:VDAC1 protein, human; D050995:Voltage-Dependent Anion Channel 1