title: Human dermal fibroblasts synthesize laminin.
abstract: Laminin, a glycoprotein of approximately 900,000 daltons, is a major component of the basement membrane that separates the epidermis from dermis in human skin. Previous studies have shown that keratinocytes and other epithelial cells synthesize laminin and utilize it for attachment to other extracellular matrices such as heparan sulfate proteoglycan and basement membrane collagen. The relationships between phenotypically normal mesenchymal cells and laminin have been much less emphasized in the literature. In this study, we have used antibodies that specifically label the A and B chains of laminin (but not fibronectin or other unrelated proteins) by Western blot analysis to immunoprecipitate biosynthetically derived laminin from [35S] methionine labeled cultures of neonatal and adult human skin fibroblasts. To be sure that the precipitated bands were laminin and not fibronectin, which has a molecular size very close to that of the laminin B chains, experiments were performed in which fibronectin was removed from the radiolabeled proteins by first immunoprecipitating with antifibronectin antibody and then sequentially immunoprecipitating laminin from the fibronectin-depleted supernates with antilaminin antibody. These experiments definitively demonstrate that human dermal fibroblasts synthesize and secrete laminin.
authors.affiliation: Department of Dermatology, UNC School of Medicine, Chapel Hill.
authors.forename: D T
authors.initials: DT
authors.lastname: Woodley
authors.forename: J R
authors.initials: JR
authors.lastname: Stanley
authors.forename: M J
authors.initials: MJ
authors.lastname: Reese
authors.forename: E J
authors.initials: EJ
authors.lastname: O'Keefe
mesh_terms: D000328:Adult; D005347:Fibroblasts / Q000032:analysis / Q000378:metabolism; D005353:Fibronectins / Q000032:analysis; D006801:Humans; D007158:Immunologic Techniques; D007231:Infant, Newborn; D007797:Laminin / Q000032:analysis / Q000096:biosynthesis*; D008970:Molecular Weight; D012867:Skin / Q000166:cytology / Q000378:metabolism*
chemical_list: D005353:Fibronectins; D007797:Laminin