Hydrophobic Chromatography in the Purification of the Histidine-binding Protein J from Salmonella typhimurium
Archives of Biochemistry and Biophysics
Upon increasing the length of the side chains in a homologous series of ω-aminoalkyl agaroses (Seph—Cn—NH2), more and more proteins become adsorbed onto the column and it is thus possible to achieve purification by selective exclusion of a desired protein. This principle is illustrated in the purification of the histidine-binding protein j from Salmonella typhimurium, vising ω-aminodecyl or ω-aminododecyl agarose columns. The protein was purified 7- to 10-fold in one step from either the shock fluid or the crude extract of the bacteria.
Since protein j has an isoelectric pH of 5.5 and yet does not bind to the positively charged columns at pH 7, it seems that this protein is not likely to have available hydrophobic pockets and may thus be remarkably hydrophilic, compared with the other proteins in the shock fluid.