Purification of interferon gamma-interferon gamma receptor complexes by preparative electrophoresis on native gels.
Cytokine-receptor complexes are required for certain studies including crystallization, NMR spectra, and investigation of the biological response mechanism to the cytokine. The purity of the ligand-receptor complex is critical for most of these applications. We investigated the possibility of purifying protein-protein complexes by electrophoresis on native gels. Starting with partially purified mouse and highly purified human proteins, we prepared milligram amounts of interferon gamma-interferon gamma receptor complexes by preparative electrophoresis on nondenaturing polyacrylamide gels. In both cases, pure ligand-receptor complexes with the correct stoichiometry of binding were recovered. Electrophoresis on preparative native gels may prove to be of general interest for the preparation of protein-protein complexes to be used in diverse studies.
Fountoulakis
M
M
F. Hoffmann-La Roche Ltd, Pharmaceutical Research-New Technologies, Basel, Switzerland.
Takacs-di Lorenzo
E
E
Juranville
J F
JF
Manneberg
M
M
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005069:Evaluation Studies as Topic; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D011485:Protein Binding; D011487:Protein Conformation; D017471:Receptors, Interferon / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D000096743:Interferon gamma Receptor
D000596:Amino Acids; D007371:Interferon-gamma; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D000096743:Interferon gamma Receptor