5-Oxoprolinase from rat kidney, II. Molecular weight determinations

The molecular weight of 5-oxoprolinase from rat kidney was estimated by gel filtration on Sephadex G-200 and G-150 to be 460 000 +/- 30 000. A value of 230 000 +/- 10 000 was obtained by zonal sedimentation in a sucrose gradient. Polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate yielded a molecular weightof 115 000 +/- 6 000. It is concluded that 5-oxoprolinase consists of four subunits of 115 000 daltons each. The dissociation or aggregation behavior of the enzyme seems to be influenced neither by the presence of the substrates 5-oxo-L-proline and MgATP2theta nor by the presence of the stabilizing compounds glutathione mercaptoethanol or dithioerythritol.