One-step chromatographic purification procedure of a His-tag recombinant carboxyl half part of the HTLV-I surface envelope glycoprotein overexpressedin Escherichia coli as a secreted form

A His-tag recombinant carboxyl half part of the HTLV-I surface envelope gly coprotein was overexpressed in E. coli as a secreted form in order to study its biochemical properties and to determine its three-dimensional structur e by X-ray crystallography. Starting from several hundred milliliters of cu lture, a centrifugation was used to eliminate the cells. After solubilizati on and centrifugation, the protein was then purified by a one-step chromato graphic purification procedure. Immobilized Metal Affinity Chromatography ( IMAC) was performed by evaluating the tri-dentate iminodiacetic acid (IDA) chelating group with chelating Sepharose fast flow, and the tetra-dendate n itrilotriacetic acid (NTA) chelating group with NTA-agarose. The latter was the most suitable gel for our protein. This expression system and the use of affinity chromatography is a rapid technique to obtain a soluble protein for use in structural studies to further understand the mechanisms of HTLV -1 entry into target cells. (C) 2001 Elsevier Science B.V. All rights reser ved.