Open AccessPurification of the β subunit of the fibronectin receptor
Author(s) -
TARONE Guido,
MASCARELLO Patrizia,
ZIBETTI Maurizio,
GIANCOTTI Filippo G.
Publication year - 1988
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1988.tb13947.x
Subject(s) - chromatography , elution , wheat germ agglutinin , chemistry , affinity chromatography , sepharose , antiserum , agarose , glycoprotein , sodium , fibronectin , lectin , sodium dodecyl sulfate , biochemistry , biology , cell , enzyme , antigen , organic chemistry , genetics
In this work we describe a method for purification of the β subunit of the mouse fibronectin receptor (CP135). Cellular glycoproteins were isolated from a detergent extract of SR‐Balb tumor cell membranes by two steps of affinity chromatography on lentil lectin‐Sepharose and wheat‐germ‐agglutinin–agarose. This material was subsequently bound to an Affi gel 102 column and eluted by increasing salt concentration. Most of the GP135 was eluted at 80 mM sodium chloride together with a few other components. A final step of hydroxyapatite chromatography in sodium dodecyl sulphate allowed elution of GP135 as a single chromatographic peak. Fractions containing GP135 were identified at each chromatographic step by immunoblotting with a specific antiserum. By this procedure GP135 was purified to homogeneity as judged by SDS‐PAGE analysis of 125 I‐labelled material.