Premium
Structure, heterologous expression, and adhesive properties of the P 0 ‐like myelin glycoprotein IP1 of trout CNS
Author(s) -
Lanwert C.,
Jeserich G.
Publication year - 2001
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.1048
Subject(s) - transfection , microbiology and biotechnology , heterologous , trout , chinese hamster ovary cell , western blot , recombinant dna , glycoprotein , chemistry , fusion protein , biology , biochemistry , gene , fishery , fish <actinopterygii> , receptor
The IP1 protein of trout CNS myelin as well as an IP1/P 0 chimeric protein were stably expressed in CHO cells. Successful targeting of the recombinant proteins to the membrane surface was verified by immunofluorescence staining. Full‐length expression of IP1 could be confirmed by Western blot analysis of proteins extracted from stably transfected CHO‐cells. The adhesive properties of IP1 were studied by an in vitro aggregation assay in which microscopic examination was combined with electronic particle counting. While IP1 conveyed only a weak increase in cell aggregation of transfected CHO cells, the IP1/P0 chimera was much more effective. In the presence of specific antibodies, cell aggregation was strongly reduced. The adhesive properties of P 0 ‐like proteins are discussed considering recent crystallographic data on the atomic structure of the extracellular domain of mammalian P 0 . Microsc. Res. Tech. 52:637–644, 2001. © 2001 Wiley‐Liss, Inc.