SEPARATION OF THE 35000 Mr DCCD-REACTIVE PROTEIN FROM THE PHOSPHATE CARRIER AND ITS PURIFICATION FROM HEART MITOCHONDRIA

Publisher Summary The C-dicyclohexylcarbodiimide (DCCD)-reactive protein of Mr more accurately determined of 35 k can be separated from the phosphate carrier and describes a procedure for its purification based on the use of an organomercurial agarose column. To separate the 35K DCCD-reactive protein from the Pi carrier, the organomercurial agarose gel, recently used to further purify the Pi carrier from the hydroxylapatite (HTP) eluate, was employed. About half of the proteins present in the HTP eluate pass through the organomercurial agarose column, whereas the remaining part is retarded and can be eluted by the addition of mercaptoethanol. The entire DCCD radioactivity is found in the first protein peak, whereas the Pi transport activity is retarded. This result definitely shows that the 35K DCCD-reactive protein and the Pi carrier are different proteins. Further purification of the DCCD-reactive protein is achieved by applying the pass-through of the organomercurial agarose column on a celite column. The reported procedure organomercurial agarose results in a pure preparation of the 35K DCCD-reactive protein