Direct visualization of affected collagen molecules synthesized by cultured fibroblasts from an osteogenesis imperfecta patient

Biochem Biophys Res Commun. 1990 Oct 15;172(1):217-22. doi: 10.1016/s0006-291x(05)80196-2.

Abstract

Human skin fibroblasts obtained from normal controls and a patient with osteogenesis imperfecta were cultured in the presence of ascorbic acid 2-phosphate, a long-acting vitamin C derivative. Crude collagen samples extracted from the cell layer were made to form lateral aggregates of collagen molecules, segment-long-spacing crystallites. Under the electron microscope, normal and abnormal crystallites of type I collagen were identified with the patient's collagen. While the carboxyl-terminal half of the abnormal crystallite was tightly packed, the amino-terminal half was loose and spreading, indicating the site of abnormality in the amino-terminal half of one of type I collagen alpha chains. The method is simple and useful to detect abnormal collagen and to predict the site of mutation.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Amino Acid Sequence
  • Cells, Cultured
  • Collagen / biosynthesis
  • Collagen / ultrastructure*
  • Female
  • Fibroblasts / metabolism
  • Humans
  • Infant, Newborn
  • Male
  • Microscopy, Electron
  • Mutation
  • Osteogenesis Imperfecta / metabolism*
  • Protein Conformation
  • Reference Values
  • Skin / metabolism*

Substances

  • Collagen