title: The amino acid tryptophan prevents the biosynthesis of dermatan sulfate.
abstract: An important part of the biosynthesis of proteoglycans is the epimerization of glycosaminoglycan chains. As a consequence of the conversion of chondroitin sulfate (CS) to dermatan sulfate (DS), the glycosaminoglycans become more flexible and enable DS to perform more sophisticated signaling functions. In a recent study, we generated a chimera (S222A) composed of a truncated form of a DS (decorin) and CS (CSF-1) containing proteoglycan and analyzed the influence of the core protein on the extent of epimerization. C-terminal truncation constructs from S222A enabled us to identify an amino acid segment that lies within the CSF-1 part which prevents DS synthesis. Co-localization experiments using S222A-HA and DCN-Flag showed different intracellular localizations for the proteoglycans during biosynthesis. A data base search revealed a sequence motif (TNWVP) within the CSF-1 moiety that is found to be important in other proteoglycans. A single substitution of tryptophan-216 to leucine (W216L) in the chimera S222A increased the amount of l-IdoA to 12-16%. Co-localization with an ER-marker demonstrated that the biosynthesis of recombinant decorin is similar to the chimera S222A and S222A(W216L) in HEK293 cells. Co-staining of S222A-HA and S222A(W216L)-Flag showed different intracellular localizations for the proteoglycans. A more detailed analysis of the glycosaminoglycans reflects a similar total sulfate content for S222A and S222A(W216L). The 4/6 sulfation ratio was similar for decorin and S222A, but altered for S222A(W216L). However, the binding of fibroblasts growth factor-1 to CS/DS was only partially dependent on epimerization. These results are consistent with the model in which the core protein, via the amino acid tryptophan, is responsible for routing to subcellular compartments with or without sufficient access to chondroitin-glucuronate 5-epimerase.
authors.affiliation: Institute of Physiological Chemistry and Pathobiochemistry, University Hospital Münster, 48149 Münster, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Herzog
authors.forename: Ines
authors.initials: I
authors.lastname: Lippmann
authors.forename: Kay
authors.initials: K
authors.lastname: Grobe
authors.forename: Alina D
authors.initials: AD
authors.lastname: Zamfir
authors.forename: Frank
authors.initials: F
authors.lastname: Echtermeyer
authors.forename: Daniela G
authors.initials: DG
authors.lastname: Seidler
mesh_terms: D001692:Biological Transport; D002460:Cell Line; D003871:Dermatan Sulfate / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D014364:Tryptophan / Q000494:pharmacology*
chemical_list: D003871:Dermatan Sulfate; D014364:Tryptophan