Expression, characterization and purification of simian immunodeficiency virus soluble, oligomerized gp160 from mammalian cells

• Published in: Journal of general virology Vol. 75; no. 1; pp. 207 - 213
• Main Authors: Rhodes, Andrew D, Spitali, Mariangela, Hutchinson, Gillian, Rud, Erling W, Stephens, Paul E
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.01.1994; Society for General Microbiology
• Subjects: AIDS/HIV; Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; CHO Cells; Cricetinae; Fundamental and applied biological sciences. Psychology; Gene Products, env - chemistry; Genetic Vectors; HIV Envelope Protein gp160; Macaca mulatta; Microbiology; Miscellaneous; Molecular Sequence Data; Protein Precursors - chemistry; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Retroviridae Proteins - biosynthesis; Retroviridae Proteins - chemistry; Retroviridae Proteins - isolation & purification; simian immunodeficiency virus; Simian Immunodeficiency Virus - genetics; Simian Immunodeficiency Virus - metabolism; Solubility; Viral Envelope Proteins - biosynthesis; Viral Envelope Proteins - chemistry; Viral Envelope Proteins - isolation & purification; Virology; Virus; Purification; Retroviridae; Lentivirinae; Biosynthesis; Glycoproteins; Oligomer; Simian immunodeficiency virus; Virus envelope; Precursor
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-75-1-207

1 Celltech Ltd, 216 Bath Road, Slough, Berkshire SL1 4EN and The 2 Wellcome Foundation, Department of Molecular Sciences, Wellcome Research Laboratories, Langley Court, Beckenham, Kent BR3 3BS, U.K. 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. Present address: The Wellcome Foundation, Department of Biochemical Sciences, Wellcome Research Laboratories, Langley Court, Beckenham, Kent BR3 3BS, U.K. Received 23 June 1993; accepted 8 September 1993.