CALCIUM-BINDING TO SIALIC ACIDS AND ITS EFFECT ON THE CONFORMATION OF EPENDYMINS

Soluble ependymins form the predominant protein constituents in the cerebrospinal fluid from many orders of teleost fish. Furthermore, these glycoproteins also exist in a bound form associated with the extracellular matrix. Ependymins are synthesized in meningeal fibroblasts. In goldfish, their synthesis is increased during the regeneration of the optic nerve and they share several characteristics with molecules involved in cell contact phenomena. In this study, we show by a calcium overlay technique that ependymins from goldfish and rainbow trout are able to bind Ca-45(2+). However, nearly all of this Ca2+-binding capacity is lost after digestion with sialidase. Furthermore, circular-dichroism spectra from FPLC-purified rainbow trout ependymins have been recorded in the presence and absence of Ca2+. Below 250 nm, the CD spectrum showed a characteristic minimum of ellipticity at 217 nm typical of beta structures. This signal is independent of the Ca2+ concentration. In contrast, the complex signal at 250-310 nm mainly decreased with increasing Ca2+ concentration indicating changes in the environment of aromatic side chains.