A study of collagen metabolism in cell cultures by fluorometric determination of proline and hydroxyproline. 1985

G Bellon, and A Randoux, and J P Borel

A technique of derivatization of proline (Pro) and 4-hydroxyproline (Hyp) by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole permitted the measurement of Pro and Hyp radioactivities, concentrations, and specific activities in the main fractions separated from cultures of fibroblast cells (extracellular collagen and non-collagen proteins, intracellular free Pro and Hyp, Pro- and Hyp-containing peptides, procollagen, and non-collagen proteins). The evaluation of collagen in the medium was obtained from as few as 10(4) cells. The method might advantageously replace [14C] Pro or [3H] Pro incorporation studies. It permits measurement of the size of the Pro pool and the amount of peptides formed by intracellular catabolism of collagen. It demonstrates that the time necessary for a full equilibration of intracellular Pro and intracellular collagen is longer than is generally believed. It avoids the uncertainties of protein labelling, which may vary with uncontrolled variations of the intracellular Pro specific activity.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008297 Male Males
D009327 4-Chloro-7-nitrobenzofurazan A benzofuran derivative used as a protein reagent since the terminal N-NBD-protein conjugate possesses interesting fluorescence and spectral properties. It has also been used as a covalent inhibitor of both beef heart mitochondrial ATPase and bacterial ATPase. Chloronitrobenzoxadiazole,NBD Chloride,7-Chloro-4-nitrobenzofurazan,NBF-Cl,Nitrobenzoxadiazole Chloride,4 Chloro 7 nitrobenzofurazan,7 Chloro 4 nitrobenzofurazan,Chloride, NBD,Chloride, Nitrobenzoxadiazole,NBF Cl
D011392 Proline A non-essential amino acid that is synthesized from GLUTAMIC ACID. It is an essential component of COLLAGEN and is important for proper functioning of joints and tendons. L-Proline,L Proline
D011865 Radioisotope Dilution Technique Method for assessing flow through a system by injection of a known quantity of radionuclide into the system and monitoring its concentration over time at a specific point in the system. (From Dorland, 28th ed) Radioisotope Dilution Technic,Dilution Technic, Radioisotope,Dilution Technics, Radioisotope,Dilution Technique, Radioisotope,Dilution Techniques, Radioisotope,Radioisotope Dilution Technics,Radioisotope Dilution Techniques,Technic, Radioisotope Dilution,Technics, Radioisotope Dilution,Technique, Radioisotope Dilution,Techniques, Radioisotope Dilution
D002250 Carbon Radioisotopes Unstable isotopes of carbon that decay or disintegrate emitting radiation. C atoms with atomic weights 10, 11, and 14-16 are radioactive carbon isotopes. Radioisotopes, Carbon
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D003094 Collagen A polypeptide substance comprising about one third of the total protein in mammalian organisms. It is the main constituent of SKIN; CONNECTIVE TISSUE; and the organic substance of bones (BONE AND BONES) and teeth (TOOTH). Avicon,Avitene,Collagen Felt,Collagen Fleece,Collagenfleece,Collastat,Dermodress,Microfibril Collagen Hemostat,Pangen,Zyderm,alpha-Collagen,Collagen Hemostat, Microfibril,alpha Collagen
D005347 Fibroblasts Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules. Fibroblast
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic

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