title: Lysine hydroxylation of collagen in a fibroblast cell culture system.
abstract: The lysine (Lys) hydroxylation pattern of type I collagen produced by human fibroblasts in culture was analyzed and compared. Fibroblasts were cultured from normal human skin (NSF), keloid (KDF), fetal skin (FDF), and skin tissues of Ehlers-Danlos syndrome type VIA and VIB patients (EDS-VIA and -VIB). The type I collagen alpha chains with or without non-helical telopeptides were purified from the insoluble matrix and analyzed. In comparison with NSFs, KDF and FDF showed significantly higher Lys hydroxylation, particularly in the telopeptide domains of both alpha chains. Both EDS-VIA and -VIB showed markedly lower Lys hydroxylation in the helical domains of both alpha chains whereas that in the telopeptides was comparable with those of NSFs. A similar profile was observed in the tissue sample of the EDS-VIB patient. These results demonstrate that the Lys hydroxylation pattern is domain-specific within the collagen molecule and that this method is useful to characterize the cell phenotypes in normal/pathological connective tissues.
authors.affiliation: Dental Research Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7455, USA.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Uzawa
authors.forename: Heather N
authors.initials: HN
authors.lastname: Yeowell
authors.forename: Kazushi
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Mochida
authors.forename: Hideki
authors.initials: H
authors.lastname: Tanzawa
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Yamauchi
mesh_terms: D000328:Adult; D002478:Cells, Cultured; D002675:Child, Preschool; D024042:Collagen Type I / Q000737:chemistry* / Q000378:metabolism*; D004535:Ehlers-Danlos Syndrome / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D008239:Lysine / Q000378:metabolism*; D008297:Male; D017434:Protein Structure, Tertiary; D012867:Skin / Q000166:cytology
chemical_list: D024042:Collagen Type I; D008239:Lysine