Incorporation of D-[6- 3H]glucosamine into maturing bone marrow granulocytes

• Published in: Biochimica et biophysica acta Vol. 273; no. 1; pp. 157 - 164
• Main Authors: Huang, Freesia L., Evans, Warren H.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 26.06.1972
• Subjects: Acetates - biosynthesis; Animals; Autoradiography; Bone Marrow - drug effects; Bone Marrow - growth & development; Bone Marrow - metabolism; Bone Marrow Cells; Centrifugation, Density Gradient; Chromatography, Paper; Glucosamine - biosynthesis; Glucosamine - metabolism; Glycoproteins - biosynthesis; Guinea Pigs; Kinetics; Leucine - metabolism; Male; Nucleoside Diphosphate Sugars - biosynthesis; Protein Biosynthesis; Puromycin - pharmacology; Solubility; Time Factors; Trichloroacetic Acid; Tritium
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/0304-4165(72)90203-6

1. 1. The incorporation of D-[6- 3H]glucosamine into cellular macromolecules was studied in guinea pig bone marrow granulocytes. 2. 2. Autoradiographic studies of labeled cells and studies on the incorporation of tritiated glucosamine into the trichloroacetic acid-insoluble fraction of granulocytes separated from other bone marrow cells by velocity sedimentation at unit gravity showed that mature granulocytes (polymorphonuclears + bands) incorporate glucosamine at a much greater rate than immature cells (myelocytes + promyelocytes + blasts). In contrast to these findings, autoradiographic studies of L-[4,5- 3H 2]leucine uptake by cells, indicated that total protein synthesis was much greater in immature than mature granulocytes. 3. 3. The major labeled compound in the hydrolysate of the trichloroacetic acid-insoluble fraction corresponded to hexosamine standard on paper chromatograms. Labeled UDP- N-acetylglucosamine and N-acetylglucosamine accounted for 31 and 25%, respectively, of the total counts in the acid-soluble pool. The incorporation of labeled glucosamine into the acid-insoluble fraction was inhibited 60% by puromycin (1.0 mM) added simultaneously with the isotope. The inhibition was significantly greater (76%) when the inhibitor was added 2 h before the addition of isotope. 4. 4. The data suggest that glucosamine is incorporated into glycoprotein of bone marrow granulocytes during the later stages of cell maturation.