Expression and biological activity of genetic fusions between MalE, the maltose binding protein from Escherichia coli and portions of CD4, the T-cell receptor of the AIDS virus.
Hybrid molecules between MalE, the periplasmic maltose binding protein of Escherichia coli, and CD4, the human T-lymphocyte receptor for the AIDS virus HIV, have been constructed and purified. We show that CD4 can be fused as multiple repeats to both ends of a single MalE molecule. Hybrid proteins are exported into the periplasm of bacteria, bind monoclonal antibodies directed against CD4, bind HIV gp160, and inhibit HIV binding to CD4+ cells. MalE has been used as a scaffold to graft portions of CD4. Deletion analysis allowed to define a minimal structural domain which folds in a way which is compatible with its biological activity. This minimal part was used to design compact hybrid molecules in which CD4 was inserted internally into MalE.
Clément
J M
JM
Unité de Programmation Moléculaire et Toxicologie Génétique (CNRS UA1444), Institut Pasteur, Paris, France.
Jehanno
M
M
Popescu
O
O
Saurin
W
W
Hofnung
M
M
D018528:ATP-Binding Cassette Transporters; D000163:Acquired Immunodeficiency Syndrome / Q000821:virology; D000911:Antibodies, Monoclonal / Q000276:immunology; D015704:CD4 Antigens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D015870:Gene Expression; D019302:HIV Envelope Protein gp160 / Q000737:chemistry; D015497:HIV-1 / Q000187:drug effects; D006801:Humans; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D010957:Plasmids / Q000235:genetics; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000821:virology
D018528:ATP-Binding Cassette Transporters; D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D019302:HIV Envelope Protein gp160; C461619:MalE protein, E coli; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; C461599:maltose transport system, E coli