Separation of Membrane Proteins of H. halobium by Gel Permeation High Performance Liquid Chromatography

概要

論文の詳細を見る
High performance liquid chromatography (HPLC) separation of the SDS-solubilized membrane proteins of Halobacterium halobium S9 was investigated with a gel permeation column, TSK G3000 or 4000 SW. When sodium phosphate buffer (pH 7.0) containing 0.1% SDS was used for elution, both the elution volume and the elution profile of protein peaks were highly dependent on the buffer concentration. On increasing the buffer concentration, the elution volume for each increased and the resolution was also improved. Protein was recovered quantitatively from the column in the buffer concentration range between 0.01 to 0.2 M. In contrast to the phosphate buffer system, the elution profile of the membrane protein was almost independent of the salt concentration when Tris-HCl (pH 7.0) buffer containing 0.1% SDS and various amounts of NaCl was used for elution. The best separation conditions finally determined for the membrane proteins of H. halobium using SW columns were as follows : 0.2 M Tris-HCl (pH 7.0) containing 0.1 M NaCl and 0.1% SDS as an elution buffer system, at a flow rate of 0.6 ml per min. Under the above elution conditions, nine major protein peaks could be determined in the SDS-solubilized plasma membrane preparation of H. halobium S9 within 40 min when the absorbance was recorded at 280 nm. The resolution obtained is comparable to that of SDS-PAGE.
社団法人日本薬学会の論文
1982-11-25