PMID: 9555098May 23, 1998Paper

Characterization of hyaluronan synthase from a human glioma cell line

Biochimica Et Biophysica Acta
T AsplundParaskevi Heldin

Abstract

In the present study we describe a method to prepare membranes with high hyaluronan synthase activity from human glioma cells by pretreatment of the cells with both testicular hyaluronidase and 4-phorbol 12-myristate 13-acetate (PMA). A 23-fold increase in hyaluronan synthase activity was detected in comparison to untreated cells. Using isolated membranes as a source of hyaluronan synthase activity we demonstrate that chain elongation occurs at the reducing end of the hyaluronan molecule. We also present a method to solubilize hyaluronan synthase in active form with 1% digitonin. The solubilized synthase synthesized shorter hyaluronan chains than the membrane bound enzyme. Partial purification of the solubilized enzyme on a Superdex-200 column revealed a 12-fold increase in specific activity. Affinity purified polyclonal antibodies, raised against a synthetic peptide corresponding to the carboxy-terminus of the deduced protein sequence of human hyaluronan synthase recognized a 66 kDa component in the purified preparations. The elution position of the solubilized hyaluronan synthesizing activity immediately after V0 corresponding to a molecular mass of about 600 kDa, suggested that the 66 kDa enzyme forms a complex with other co...Continue Reading

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Citations

Jul 14, 2004·Biochemistry·Valarie L Tlapak-SimmonsPaul H Weigel
May 5, 2000·The Biochemical Journal·A JacobsonP Heldin
May 29, 2010·The Journal of Biological Chemistry·Eugenia KarousouParaskevi Heldin
Jan 26, 2005·The Journal of Biological Chemistry·Valarie L Tlapak-SimmonsPaul H Weigel
May 13, 2008·Clinics in Dermatology·Robert Stern, Howard I Maibach
May 27, 2003·The Journal of Physiology·C L AnggiansahJ R Levick
Jan 19, 2008·The Journal of Physiology·K R IngramJ R Levick
Oct 19, 2013·Matrix Biology : Journal of the International Society for Matrix Biology·Davide VigettiAlberto Passi
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Jan 15, 2015·Glycobiology·Paul H WeigelJennifer L Washburn
Apr 4, 2003·Biological Chemistry·Johannes MülleggerGünter Lepperdinger
Mar 26, 2003·The Journal of Biological Chemistry·Philip E Pummill, Paul L DeAngelis
Nov 6, 2007·The Journal of Biological Chemistry·Paul H Weigel, Paul L DeAngelis
Oct 30, 2003·The Journal of Biological Chemistry·Hiroko HoshiShin-Ichiro Nishimura
May 25, 2006·The Biochemical Journal·Peter Prehm
Dec 30, 2004·The Journal of Biological Chemistry·Sabrina Bodevin-AutheletUlf Lindahl
Mar 22, 2021·The Journal of Biological Chemistry·Matías Arturo PibuelSilvina Laura Lompardía

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