Structure and function of the carboxyl-terminal oxygen-binding domain from the subunit of Octopus dofleini hemocyanin.
The C-terminal domain, Od-1, of the 7-domain subunit of Octopus dofleini hemocyanin has been prepared by partial trypsinolysis followed by ion-exchange chromatography. It binds oxygen reversibly and is homogeneous in molecular weight. Its physical properties have been compared with those of the subunit. The domain molecular weight is found by sedimentation equilibrium to be 4.7 X 10(4), in excellent agreement with the result recently obtained in our laboratory from cDNA sequencing of this domain [Lang, W. H. (1988) Biochemistry (preceding paper in this issue)]. It has a sedimentation coefficient of 3.8 S. Both the molecular weight and sedimentation coefficient are consistent with the domain constituting approximately one-seventh of the Mr 3.5 X 10(5) subunit. Its amino acid composition and carbohydrate content differ significantly from that of the whole subunit, confirming the heterogeneity in domains previously established on an immunological basis. Circular dichroism predicts similar secondary structure for the domain and subunit. The domain does not self-associate in the presence of Mg2+ but does bind to the whole molecule in a ratio of approximately 1 domain/subunit. The oxygen affinity of this domain is quite low. It shows intrinsic magnesium and Bohr effects similar to those of the whole molecule but of greatly reduced magnitude.
Miller
K I
KI
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331.
van Holde
K E
KE
Toumadje
A
A
Johnson
W C
WC
Lamy
J
J
D000116:Acetylgalactosamine / Q000032:analysis; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002241:Carbohydrates / Q000032:analysis; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D006433:Hemocyanins / Q000032:analysis / Q000378:metabolism*; D007122:Immunoelectrophoresis; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D049831:Octopodiformes / Q000032:analysis*; D010100:Oxygen / Q000378:metabolism*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D014357:Trypsin / Q000378:metabolism; D014461:Ultracentrifugation
D000596:Amino Acids; D002241:Carbohydrates; D006433:Hemocyanins; D014357:Trypsin; D008274:Magnesium; D000116:Acetylgalactosamine; D010100:Oxygen