Purification of the beta subunit of the fibronectin receptor.
In this work we describe a method for purification of the beta subunit of the mouse fibronectin receptor (GP135). 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 125I-labelled material.
Tarone
G
G
Department of Biomedical Sciences and Human Oncology, University of Torino, Italy.
Mascarello
P
P
Zibetti
M
M
Giancotti
F G
FG
D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D002846:Chromatography, Affinity; D005354:Fibrosarcoma / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic / Q000032:analysis*
D006023:Glycoproteins; D008565:Membrane Proteins; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic