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The extracellular domain of immunodeficiency virus gp41 protein: Expression in Escherichia coli , purification, and crystallization
Author(s) -
Wingfield Paul T.,
Stahl Stephen J.,
Kaufman Joshua,
Zlotnick Adam,
Hyde C. Craig,
Gronenborn Angela M.,
Clore G. Marius
Publication year - 1997
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560060806
Subject(s) - escherichia coli , gp41 , crystallization , extracellular , human immunodeficiency virus (hiv) , virology , protein expression , biology , domain (mathematical analysis) , microbiology and biotechnology , chemistry , biochemistry , genetics , antigen , gene , epitope , organic chemistry , mathematical analysis , mathematics
The env gene of SIV and HIV‐1 encodes a single glycoprotein gp160, which is processed to give a noncovalent complex of the soluble glycoprotein gp120 and the transmembrane glycoprotein gp41. The extracellular region (ectodomain), minus the N‐terminal fusion peptide, of gp41 from HIV‐1 (residues 27‐154) and SIV (residues 27‐149) have been expressed in Escherichia coli . These insoluble proteins were solubilized and subjected to a simple purification and folding scheme, which results in high yields of soluble protein. Purified proteins have a trimeric subunit composition and high a‐helical content, consistent with the predicted coil‐coil structure. SIV gp41 containing a double cysteine mutation was crystallized. The crystals are suitable for X‐ray structure determination and, preliminary analysis, together with additional biochemical evidence, indicates that the gp41 trimer is arranged as a parallel bundle with threefold symmetry.