Production of secreted, soluble human two-domain CD4 protein in Escherichia coli.
The two-domain form of recombinant soluble human CD4 (rsCD4(183)) has been used for structural studies and to probe the interaction of CD4 with its ligands. rsCD4(183) has generally been produced in Escherichia coli in the form of inclusion bodies. The generation of conformationally native protein from these inclusion bodies is a time-consuming and inefficient process, requiring a refolding step. Here, we describe a procedure for producing 2-4 mg of secreted, conformationally native rsCD4(183) per liter of E. coli, completely bypassing the requirement for protein refolding in vitro. Furthermore, the yield of active protein is comparable to that reported for expression systems that generate inclusion bodies.
Osburne
M S
MS
Procept, Cambridge, MA 02139, USA. osburnem@ariad.com
Neidhardt
E A
EA
Godoy
J E
JE
van Schravendijk
M R
MR
Grossman
T H
TH
D015704:CD4 Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D002457:Cell Extracts; D004926:Escherichia coli; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012995:Solubility
D015704:CD4 Antigens; D002457:Cell Extracts; D011993:Recombinant Fusion Proteins