title: Modulation of complement regulatory function and measles virus receptor function by the serine-threonine-rich domains of membrane cofactor protein (CD46).
abstract: Three major membrane cofactor protein (MCP) phenotypes with different serine-threonine (ST)-rich regions, namely STc (L-phenotype), STBC (H or U phenotype) and STABC, and the MCP without the ST domain (delta ST) were expressed in Chinese hamster ovary (CHO) cells by transfecting the respective cDNAs. The expressed molecules migrated with a larger molecular mass on SDS/PAGE than those expected from their amino acid sequences. O-Glycanase digestion showed that this was due to O-linked sugar chains. The apparent sugar contents in each ST segment were compatible with their serine and threonine contents in the ST regions. The functional properties of these phenotypes as inhibitors of human complement (C) and receptors of measles virus (MV) were compared. The classical pathway-dependent CHO cell lysis by human C was more effectively suppressed by the expressed delta ST and STC than by the STABC and STBC phenotypes, although the difference was not so prominent. In contrast, alternative C pathway-dependent CHO-cell lysis was most effectively suppressed by the STABC phenotype and was only slightly blocked by the ST-deleted mutant. MV infection occurred with all of the phenotypes, but the infectious dose required to cause the same level of syncytium formation was 100-times higher in large ST (STABC and STBC) than in small ST (STC and delta ST) phenotypes. Thus, the ST domain serves as a functional modulator in MCP: MCP with a large ST domain having high O-linked sugar contents is favourable to the effective suppression of both the alternative C pathway-mediated cytolysis and MV infection, whereas MCP with a small ST domain is favourable to the suppression of the classical C pathway.
authors.affiliation: Department of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Iwata
authors.forename: T
authors.initials: T
authors.lastname: Seya
authors.forename: S
authors.initials: S
authors.lastname: Ueda
authors.forename: H
authors.initials: H
authors.lastname: Ariga
authors.forename: S
authors.initials: S
authors.lastname: Nagasawa
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D001483:Base Sequence; D016466:CHO Cells; D003169:Complement Inactivator Proteins / Q000378:metabolism*; D006224:Cricetinae; D018076:DNA, Complementary; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D006801:Humans; D015151:Immunoblotting; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D008969:Molecular Sequence Data; D012694:Serine / Q000737:chemistry; D013912:Threonine / Q000737:chemistry
chemical_list: D015703:Antigens, CD; C497857:CD46 protein, human; D003169:Complement Inactivator Proteins; D018076:DNA, Complementary; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D013912:Threonine; D012694:Serine