title: Divergent secretory behavior of the opposite ends of aggrecan.
abstract: The proteoglycan, aggrecan has a globular domain, G1, at the N terminus and a different globular domain, G3, at the C terminus. Aggrecan produced by mutant nanomelic chickens is truncated due to a premature stop codon and consequently lacks G3 and a minor portion of its chondroitin sulfate domain (Li, H., Schwartz, N. B., and Vertel, B. M.(1993) J. Biol. Chem. 268, 23504-23511). The mutant protein is retained in the endoplasmic reticulum and fails to enter the Golgi stacks (Vertel, B. M., Walters, L. M., Grier, B., Maine, N. , and Goetinck, P. F.(1993) J. Cell Sci. 104, 939-948). The homozygous mutant is lethal because of failure of chondrogenesis and osteogenesis, while the heterozygous mutant is dwarfed. To further elucidate the pathogenetic mechanisms underlying nanomelia and to determine if G1 and G3 are themselves secreted, we expressed them in transfected host cells. Expression was performed in wild type Chinese hamster ovary (CHO) cells and in mutant CHO cells which are unable to link glycosaminoglycan (GAG) chains to core proteins. We compared: (a) secretion of expressed G1 and G3 constructs containing contiguous GAG chain consensus sites and (b) GAG chain modification of the secreted proteins. We find that: 1) G3 is 24-100 times more rapidly secreted than G1; 2) secreted G3 contains contiguous chondroitin sulfate GAG chains, while secreted G1 lacks contiguous GAG chains; 3) G3 secretion is not coupled to xylosylation of contiguous GAG chain consensus sites. These results imply that G1 and G3 intrinsically differ in passage through the cell secretory route.
authors.affiliation: Department of Biostructure and Function, School of Dental Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030-3705, USA.
authors.forename: W
authors.initials: W
authors.lastname: Luo
authors.forename: T S
authors.initials: TS
authors.lastname: Kuwada
authors.forename: L
authors.initials: L
authors.lastname: Chandrasekaran
authors.forename: J
authors.initials: J
authors.lastname: Zheng
authors.forename: M L
authors.initials: ML
authors.lastname: Tanzer
mesh_terms: D053674:Aggrecans; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D016466:CHO Cells; D002645:Chickens; D006224:Cricetinae; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D008969:Molecular Sequence Data; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism*; D014994:Xylose / Q000378:metabolism
chemical_list: D053674:Aggrecans; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D014994:Xylose