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. 1983 Apr 1;211(1):191–198. doi: 10.1042/bj2110191

Synthesis of hyaluronate in differentiated teratocarcinoma cells. Mechanism of chain growth.

P Prehm
PMCID: PMC1154343  PMID: 6870820

Abstract

Hyaluronate could be labelled in vivo with [32P]phosphate. [32P]UDP in an alpha-glycosidic linkage constituted the reducing end of membrane-bound hyaluronate. The UDP is liberated during further chain elongation, indicating that chain growth occurs at the reducing end. [3H]Uridine could be incorporated into hyaluronate during synthesis on the isolated membraneous fraction from [3H]UDP-GlcNAc and [3H]UDP-GlcA, confirming the identification of UDP as a constituent of membrane-bound hyaluronate. These results led to a model of hyaluronate chain elongation at the reducing end by alternate addition of the chains to the substrates. Membrane-bound pyrophosphatases or 5'-nucleotidase are suggested as modulators of hyaluronate synthesis.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. AXELROD J., KALCKAR H. M., MAXWELL E. S., STROMINGER J. L. Enzymatic formation of uridine diphosphoglucuronic acid. J Biol Chem. 1957 Jan;224(1):79–90. [PubMed] [Google Scholar]
  2. Abney E. R., Evans W. H., Parkhouse R. M. Location of nucleotide pyrophosphatase and alkaline phosphodiesterase activities on the lymphocyte surface membrane. Biochem J. 1976 Nov;159(2):293–299. doi: 10.1042/bj1590293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BIELESKI R. L. SEPARATION OF PHOSPHATE ESTERS BY THIN-LAYER CHROMATOGRAPHY AND ELECTROPHORESIS. Anal Biochem. 1965 Aug;12:230–234. doi: 10.1016/0003-2697(65)90086-2. [DOI] [PubMed] [Google Scholar]
  4. Bischoff E., Tran-Thi T. A., Decker K. F. Nucleotide pyrophosphatase of rat liver. A comparative study on the enzymes solubilized and purified from plasma membrane and endoplasmic reticulum. Eur J Biochem. 1975 Feb 21;51(2):353–361. doi: 10.1111/j.1432-1033.1975.tb03935.x. [DOI] [PubMed] [Google Scholar]
  5. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bordier C. Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem. 1981 Feb 25;256(4):1604–1607. [PubMed] [Google Scholar]
  7. CRANE R. K., LIPMANN F. The relationship of mitochondrial phosphate to aerobic phosphate bond generation. J Biol Chem. 1953 Mar;201(1):245–246. [PubMed] [Google Scholar]
  8. Castor C. W., Dorstewitz E. L., Rowe K., Ritchie J. C. Abnormalities of connective tissue cells cultured from patients with rheumatoid arthritis. II. Defective regulation of hyaluronate and collagen formation. J Lab Clin Med. 1971 Jan;77(1):65–75. [PubMed] [Google Scholar]
  9. Fransson L. A., Rodén L. Structure of dermatan sulfate. I. Degradation by testicular hyaluronidase. J Biol Chem. 1967 Sep 25;242(18):4161–4169. [PubMed] [Google Scholar]
  10. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hart G. W., Lennarz W. J. Effects of tunicamycin on the biosynthesis of glycosaminoglycans by embryonic chick cornea. J Biol Chem. 1978 Aug 25;253(16):5795–5801. [PubMed] [Google Scholar]
  12. Haugen H. F., Skrede S. Nucleotide pyrophosphatase and phosphodiesterase. I. Organ distribution and activities in body fluids. Clin Chem. 1977 Sep;23(9):1531–1537. [PubMed] [Google Scholar]
  13. Hopwood J. J., Dorfman A. Glycosaminoglycan synthesis by cultured human skin fibroblasts after transformation with simian virus 40. J Biol Chem. 1977 Jul 25;252(14):4777–4785. [PubMed] [Google Scholar]
  14. Ishimoto N., Strominger J. L. Uridine diphosphate as the sole uridine nucleotide product of hyaluronic acid synthetase in group A streptococci. Biochim Biophys Acta. 1967 Oct 9;148(1):296–297. doi: 10.1016/0304-4165(67)90305-4. [DOI] [PubMed] [Google Scholar]
  15. Katzman R. L. Absence of arabinose in bovine brain hyaluronic acid as analysed by gas--liquid chromatography and mass spectrometry. Biochim Biophys Acta. 1974 Nov 4;372(1):53–54. [PubMed] [Google Scholar]
  16. Kjellén L., Pettersson I., Hök M. Cell-surface heparan sulfate: an intercalated membrane proteoglycan. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5371–5375. doi: 10.1073/pnas.78.9.5371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kleine T. O. Biosynthesis of proteoglycans: an approach to locate it in different membrane systems. Int Rev Connect Tissue Res. 1981;9:27–98. doi: 10.1016/b978-0-12-363709-3.50008-5. [DOI] [PubMed] [Google Scholar]
  18. Kleine T. O. Hyaluronate-proteoglycan complex: evidence for separate biosynthesis mechanisms of the macromolecules. Connect Tissue Res. 1978;5(4):195–199. doi: 10.3109/03008207809152272. [DOI] [PubMed] [Google Scholar]
  19. Koyama H., Ono T. Further studies on the induction of alkaline phosphatase by 5-bromodeoxyuridine in a hybrid line between mouse and Chinese hamster in culture. Biochim Biophys Acta. 1972 May 16;264(3):497–507. doi: 10.1016/0304-4165(72)90013-x. [DOI] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Mankin H. J. Stimulation of hyaluronate production in joints: a worthwhile goal? J Rheumatol. 1979 Sep-Oct;6(5):481–483. [PubMed] [Google Scholar]
  22. Mapleson J. L., Buchwald M. Effect of cycloheximide and dexamethasone phosphate on hyaluronic acid synthesis and secretion in cultured human skin fibroblasts. J Cell Physiol. 1981 Nov;109(2):215–222. doi: 10.1002/jcp.1041090204. [DOI] [PubMed] [Google Scholar]
  23. Mikuni-Takagaki Y., Toole B. P. Hyaluronate-protein complex of rous sarcoma virus-transformed chick embryo fibroblasts. J Biol Chem. 1981 Aug 25;256(16):8463–8469. [PubMed] [Google Scholar]
  24. Mikuni-Takagaki Y., Toole B. P. Shedding of hyaluronate from the cell surface of Rous sarcoma virus-transformed chondrocytes. J Biol Chem. 1979 Sep 10;254(17):8409–8415. [PubMed] [Google Scholar]
  25. Mårdh S., Vega F. Kinetics of a nucleotide pyrophosphatase in the plasma membrane of the fat cell. Biochim Biophys Acta. 1980 Oct 2;601(3):524–531. doi: 10.1016/0005-2736(80)90555-6. [DOI] [PubMed] [Google Scholar]
  26. O'BRIEN P. J. THE SYNTHESIS OF N-ACETYL-ALPHA-AND N-ACETYL-BETA-D-GLUCOSAMINE I-PHOSPHATES (2-ACETAMIDO-2-DEOXY-ALPHA- AND BETA-D-GLUCOSE I-PHOSPHATES). Biochim Biophys Acta. 1964 Jun 8;86:628–634. doi: 10.1016/0304-4165(64)90103-5. [DOI] [PubMed] [Google Scholar]
  27. Prehm P. Synthesis of hyaluronate in differentiated teratocarcinoma cells. Characterization of the synthase. Biochem J. 1983 Apr 1;211(1):181–189. doi: 10.1042/bj2110181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. RANDERATH K., RANDERATH E. ION-EXCHANGE CHROMATOGRAPHY OF NUCLEOTIDES ON POLY-(ETHYLENEIMINE)-CELLULOSE THIN LAYERS. J Chromatogr. 1964 Oct;16:111–125. doi: 10.1016/s0021-9673(01)82445-6. [DOI] [PubMed] [Google Scholar]
  29. Rao A. K., Mendicino J. Synthesis of UDP-N-[1-14C]acetyl D-glucosamine and UDP-N-[1-14C]acetyl-D-galactosamine from [1-14C]acetate. Anal Biochem. 1978 Dec;91(2):490–495. doi: 10.1016/0003-2697(78)90535-3. [DOI] [PubMed] [Google Scholar]
  30. STARY Z., WARDI A. H., TURNER D. L., ALLEN W. S. ARABINOSE AS A MUCOPOLYSACCHARIDE COMPONENT IN HUMAN AND ANIMAL BRAIN TISSUE. Arch Biochem Biophys. 1965 May;110:388–394. doi: 10.1016/0003-9861(65)90139-6. [DOI] [PubMed] [Google Scholar]
  31. Sawicka T., Grzelak-Puczyńska I., Sawicki W., Bagdasarian M. Cell surface nucleotide pyrophosphatase and sugar phosphate phosphohydrolase activity: interspecies and cell type variations. Folia Histochem Cytochem (Krakow) 1979;17(4):327–334. [PubMed] [Google Scholar]
  32. Sela B. A., Lis H., Sachs L. Enzymatic hydrolysis of uridine diphosphate-N-acetyl-D-galactosamine and uridine diphosphate-N-acetyl-D-glucosamine by normal cells, and blocks in this hydrolysis in transformed cells and their revertants. J Biol Chem. 1972 Dec 10;247(23):7585–7590. [PubMed] [Google Scholar]
  33. Sugahara K., Schwartz N. B., Dorfman A. Biosynthesis of hyaluronic acid by Streptococcus. J Biol Chem. 1979 Jul 25;254(14):6252–6261. [PubMed] [Google Scholar]
  34. Swann D. A. Studies on hyaluronic acid. II. The protein component(s) of rooster comb hyaluronic acid. Biochim Biophys Acta. 1968 May 6;160(1):96–105. doi: 10.1016/0005-2795(68)90068-8. [DOI] [PubMed] [Google Scholar]
  35. Touster O., Aronson N. N., Jr, Dulaney J. T., Hendrickson H. Isolation of rat liver plasma membranes. Use of nucleotide pyrophosphatase and phosphodiesterase I as marker enzymes. J Cell Biol. 1970 Dec;47(3):604–618. doi: 10.1083/jcb.47.3.604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Varma R., Varma R. S., Allen W. S., Wardi A. H. Human umbilical cord hyaluronate. Neutral sugar content and carbohydrate-protein linkage studies. Biochim Biophys Acta. 1975 Jul 14;399(1):139–144. doi: 10.1016/0304-4165(75)90220-2. [DOI] [PubMed] [Google Scholar]
  37. Varma R., Varma R. S., Allen W. S., Wardi A. H. On the carbohydrate-protein linkage group in vitreous humor hyaluronate. Biochim Biophys Acta. 1974 Oct 8;362(3):584–588. doi: 10.1016/0304-4165(74)90154-8. [DOI] [PubMed] [Google Scholar]
  38. Varma R., Vercellotti J. R., Varma R. S. On arabinose as a component of brain hyaluronate. Confirmation by chromatographic, enzymatic and chemical ionization-mass spectrometric analyses. Biochim Biophys Acta. 1977 Apr 27;497(2):608–614. doi: 10.1016/0304-4165(77)90217-3. [DOI] [PubMed] [Google Scholar]
  39. Wardi A. H., Allen W. S., Michos G. A., Turner D. L. Isolation of unknown sugar acid from vitreous humor hyaluronic acid. Biochim Biophys Acta. 1969 Jul 30;184(2):474–476. doi: 10.1016/0304-4165(69)90055-5. [DOI] [PubMed] [Google Scholar]
  40. Winterbourne D. J., Mora P. T. Altered metabolism of heparan sulfate in simian virus 40 transformed cloned mouse cells. J Biol Chem. 1978 Jul 25;253(14):5109–5120. [PubMed] [Google Scholar]
  41. van den Berghe G., van Pottelsberghe C., Hers H. G. A kinetic study of the soluble 5'-nucleotidase of rat liver. Biochem J. 1977 Mar 15;162(3):611–616. doi: 10.1042/bj1620611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. von Figura K., Kiowski W., Buddecke E. Differently labelled glucosamine-precursor pools for the biosynthesis of hyaluronate and heparan sulfate. Eur J Biochem. 1973 Dec 3;40(1):89–94. doi: 10.1111/j.1432-1033.1973.tb03171.x. [DOI] [PubMed] [Google Scholar]

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