Expression, characterization and purification of simian immunodeficiency virus soluble, oligomerized gp160 from mammalian cells.
The envelope glycoprotein, gp160, of human (HIV) and simian (SIV) immunodeficiency viruses mediates virus-host cell binding followed by fusion of the viral and plasma membranes. The envelope proteins are known to exist as non-covalently associated oligomers on the virus surface. The production of permanent mammalian cell lines that constitutively secrete relatively high levels of soluble forms of SIV gp160 is described and we show that these proteins are secreted predominantly as tetramers with lower levels of dimer forms. Oligomeric forms were purified to greater than 90% purity using a simple gel filtration method. The purified proteins bind CD4 suggesting that they remain in their native conformation. The purified oligomeric proteins provide the basis for more relevant structural, functional and immunological studies than recombinant gp120 as they more closely resemble the envelope protein oligomer.
Rhodes
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AD
Celltech Ltd, Slough, Berkshire, U.K.
Spitali
M
M
Hutchinson
G
G
Rud
E W
EW
Stephens
P E
PE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D006224:Cricetinae; D015686:Gene Products, env / Q000737:chemistry; D005822:Genetic Vectors; D019302:HIV Envelope Protein gp160; D008253:Macaca mulatta; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012191:Retroviridae Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification
D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors; D011994:Recombinant Proteins; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins