Open AccessThe Synthesis of Macromolecular 3‐Hydroxyproline by Attaching and Confluent Cultures of Human Fibroblasts
Author(s) -
LEMBACH Kenneth J.,
BRANSON R. Edward,
HEWGLEY Paula B.,
CUNNINGHAM Leon W.
Publication year - 1977
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1977.tb11262.x
Subject(s) - hydroxyproline , trypsinization , proline , biochemistry , chemistry , substrate (aquarium) , fibroblast , macromolecule , biology , enzyme , amino acid , in vitro , trypsin , ecology
The synthesis of collagen has been studied during the attachment of freshly trypsinized human fibroblasts to culture vessels by measurement of the incorporation of radioactive proline into macro‐molecular hydroxyproline. Collagenous protein(s) was found to be a component of a substrate‐attached material (‘microexudate carpet’) synthesized rapidly during cell attachment in the absence of serum. The ratio of 3‐hydroxyproline/4‐hydroxyproline in the collagenous proteins synthesized during cell attachment was found to be 4–5‐fold higher than that of normal type I collagen. The synthesis of 3‐hydroxyproline by confluent cultures was diminished by serum deprivation, and was shown to require higher concentrations of ascorbate than the synthesis of the 4‐hydroxy isomer.