期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1990
卷号：87
期号：4
页码：1496-1500
DOI：10.1073/pnas.87.4.1496
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Native Octopus dofleini hemocyanin appears as a hollow cylinder in the electron microscope. It is composed of 10 polypeptide subunits, each folded into seven globular oxygen-binding domains. The native structure reassociates spontaneously from subunits in the presence of Mg2+ ions. We have selectively removed the C-terminal domain and purified the resulting six-domain subunits. Although these six-domain subunits do not associate efficiently at pH 7.2, they undergo nearly complete reassociation at pH 8.0. The resulting molecule looks like the native cylindrical whole molecule but lacks the usual fivefold protrusions into the central cavity. Partially reassociated mixtures show dimers of the subunit that have a characteristic parallelogram shape when lying flat on the electron microscope grid, and a "boat" form in side view. Removal of the C-terminal domain from monomers results in the removal of two characteristically placed domains in the dimers. These observations allow the development of a model for the arrangement of the subunits within the whole molecule. The model predicts exactly the views seen in the electron microscope of both whole molecule and dimeric intermediates.
