EXPORT AND ONE-STEP PURIFICATION FROM ESCHERICHIA-COLI OF A MALE-CD4 HYBRID PROTEIN THAT NEUTRALIZES HIV INVITRO
- 1 January 1990
- journal article
- research article
- Vol. 3 (9) , 859-872
Abstract
The 177 N-terminal amino acids of CD4, the receptor of the human immunodeficiency virus (HIV), have been expressed in Escherichia coli as genetic fusions to the periplasmic maltose-binding protein (MalE) from this organism. A large fraction of the hybid proteins can be released from the periplasm by osmotic shock and purified in one step on a cross-linked amylose column eluted with maltose under mild conditions. One hybrid protein binds HIV envelope protein gp160 and neutralizes the virus in vitro. This provides the first example of the production and one-step purification of an active form of an eukaryotic protein by fusion to Ma1E. The use of this system for mass screening of CD4 mutants, high-scale production of the hybrid protein for structural studies on CD4, testing antiviral compounds, and therapeutic assays is discussed.This publication has 33 references indexed in Scilit:
- Studies on transformation of Escherichia coli with plasmidsPublished by Elsevier ,2006
- Selective Tropism of Lymphadenopathy Associated Virus (LAV) for Helper-Inducer T LymphocytesScience, 1984
- Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectorsGene, 1983
- Mutations that alter the signal sequence of alkaline phosphatase in Escherichia coliJournal of Bacteriology, 1983
- Possible involvement of the OKT4 molecule in T cell recognition of class II HLA antigens. Evidence from studies of cytotoxic T lymphocytes specific for SB antigens.The Journal of Experimental Medicine, 1982
- Clonal analysis of human cytotoxic T lymphocytes: T4+ and T8+ effector T cells recognize products of different major histocompatibility complex regions.Proceedings of the National Academy of Sciences, 1982
- High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules.Proceedings of the National Academy of Sciences, 1979
- Precursors of three exported proteins in Escherichia coli.Proceedings of the National Academy of Sciences, 1978
- An efficient and reproducible procedure for the formation of spheroplasts from variously grown Escherichia coliAnalytical Biochemistry, 1976
- The Release of Enzymes from Escherichia coli by Osmotic Shock and during the Formation of SpheroplastsJournal of Biological Chemistry, 1965