Studies on erythrocruorin. VII. Reconstitution of earthworm erythrocruorin from the apoprotein.
Apoerythrocruorin prepared from the giant respiratory hemoprotein of the earthworm (60 S, Mr = 3 X 10(-6)) is an electrophoretically homogeneous molecule which sediments as a single peak of low molecular weight (3.5 S) and has a lower alpha-helical content (approx. 30%) than the native protein. Titration of globin with ferric heme indicates the presence of different binding sites; however, after purification by ion exchange chromatography, the reconstitution product contains 1 haem/23 000 g of protein as the native molecule. Reconstituted ferric erythrocruorin is a low molecular weight hemichrome with the same optical and physicochemical properties of the hemichrome formed by natural ferric erythrocruorin. Reconstituted ferrous erythrocruorin reacquires the alpha-helical content and the quaternary structure of the native molecule. Reassociation into 10-S speices (1/12 of the whole molecules) is fast and easy, while that into whole molecules is slow and somewhat erratic. The functional properties of reconstituted ferrous erythrocruorin (oxygen affinity, cooperativity in oxygen binding, magnitude of Bohr effect) are very similar to those of the "stable" low cooperativity form of the undissociated protein.
D001059:Apoproteins / Q000302:isolation & purification; D002942:Circular Dichroism; D004902:Erythrocruorins; D005290:Ferric Compounds; D005296:Ferrous Compounds; D006418:Heme; D006454:Hemoglobins; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009835:Oligochaeta; D010100:Oxygen / Q000097:blood; D011485:Protein Binding; D011487:Protein Conformation
D001059:Apoproteins; D004902:Erythrocruorins; D005290:Ferric Compounds; D005296:Ferrous Compounds; D006454:Hemoglobins; D006418:Heme; D010100:Oxygen