Detection of protein complex interactions via a Blue Native-PAGE retardation assay.
We describe the Blue Native (BN)-PAGE retardation assay for the detection of interactions of biomolecules with protein complexes. Potential interactors of proteins are included in the BN gel matrix, resulting in retardation of proteins that interact with the added molecule. After validation using the T-cell antigen receptor, we applied the assay for a general identification of dextran interactors in combination with mass spectroscopy. The proteomic screen revealed triosephosphate isomerase oligomer as a dextran-binding, high M(R) complex.
Swamy
Mahima
M
Department for Molecular Immunology, Max Planck-Institute for Immunobiology, 79108 Freiburg, Germany.
Molnar
Eszter
E
Bock
Thomas
T
Bausch-Fluck
Damaris
D
Wollscheid
Bernd
B
Schamel
Wolfgang W A
WW
D000818:Animals; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D051379:Mice; D055503:Protein Multimerization; D011948:Receptors, Antigen, T-Cell / Q000032:analysis* / Q000378:metabolism; D012394:Rosaniline Dyes
D011948:Receptors, Antigen, T-Cell; D012394:Rosaniline Dyes; C004692:coomassie Brilliant Blue