A method for the rapid and efficient elution of native affinity-purified protein A tagged complexes.
A problem faced in proteomics studies is the recovery of tagged protein complexes in their native and active form. Here we describe a peptide, Bio-Ox, that mimics the immunoglobulin G (IgG) binding interface of Staphylococcus aureus Protein A, and competitively displaces affinity-purified Protein A fusion proteins and protein complexes from IgG-Sepharose. We show that Bio-Ox elution is a robust method for the efficient and rapid recovery of native tagged proteins, and can be applied to a variety of structural genomics and proteomics studies.
Strambio-de-Castillia
Caterina
C
Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY 10021-6399, USA.
Tetenbaum-Novatt
Jaclyn
J
Imai
Brian S
BS
Chait
Brian T
BT
Rout
Michael P
MP
D001426:Bacterial Proteins / Q000737:chemistry; D001671:Biochemistry / Q000379:methods*; D001710:Biotin / Q000737:chemistry*; D002846:Chromatography, Affinity; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry*; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry; D013211:Staphylococcus aureus / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors
D001426:Bacterial Proteins; D007074:Immunoglobulin G; D010455:Peptides; D011506:Proteins; D011993:Recombinant Fusion Proteins; D001710:Biotin; D012685:Sepharose