Mutation of the sialyltransferase S-sialylmotif alters the kinetics of the donor and acceptor substrates.

The Journal of Biological Chemistry
A K DattaJ C Paulson

Abstract

Protein sequence analysis of the cloned sialyltransferase gene family has revealed the presence of two conserved protein motifs in the middle of the lumenal catalytic domain, termed L-sialylmotif and S-sialylmotif. In our previous study (Datta, A. K., and Paulson, J. C. (1995) J. Biol. Chem. 270, 1497-1500) the larger L-sialylmotif of ST6Gal I was analyzed by site-directed mutagenesis, which provided evidence that it participates in the binding of the CMP-NeuAc, a common donor substrate for all the sialyltransferases. However, none of the mutants tested in this motif had any significant effect on their binding affinities toward the acceptor substrate asialo alpha1-acid glycoprotein. In this study, we have investigated the role of the S-sialylmotif of the same enzyme ST6Gal I. In total, nine mutants have been constructed by changing the conserved amino acids of this motif to mostly alanine by site-directed mutagenesis. Kinetic analysis for the mutants which retained sialyltransferase activity showed that the mutations in the S-sialylmotif caused a change of Km values for both the donor and the acceptor substrates. Our results indicated that this motif participates in the binding of both the substrates. A sequence homology search...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Aug 1, 1990·The Journal of Experimental Medicine·I StamenkovicS Funderud
Feb 11, 1990·Nucleic Acids Research·U GrundmannG Zettlmeissl
Sep 29, 1995·The Journal of Biological Chemistry·E P ScheideggerJ B Lowe
Jul 18, 1995·Proceedings of the National Academy of Sciences of the United States of America·J NakayamaM Fukuda
May 11, 1995·Biochimica Et Biophysica Acta·N KurosawaS Tsuji
Jun 16, 1995·The Journal of Biological Chemistry·Y YoshidaS Tsuji
Jan 27, 1995·The Journal of Biological Chemistry·A K Datta, J C Paulson
Feb 23, 1995·Nature·M EckhardtR Gerardy-Schahn
Oct 25, 1994·Proceedings of the National Academy of Sciences of the United States of America·M HaraguchiK Furukawa
Oct 28, 1994·Biochemical and Biophysical Research Communications·Y WangH E Appert
Aug 16, 1994·Proceedings of the National Academy of Sciences of the United States of America·K NaraY Sanai
Jul 15, 1993·Biochemical and Biophysical Research Communications·H Kitagawa, J C Paulson
Feb 1, 1993·Glycobiology·K Drickamer
Dec 1, 1995·Glycobiology·A Harduin-LepersP Delannoy
Jul 1, 1996·Journal of Biochemistry·S Tsuji
Oct 1, 1996·Glycobiology·S TsujiJ C Paulson

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Citations

Nov 9, 2002·Medicinal Research Reviews·Xiaofang WangXin-Shan Ye
Apr 25, 2012·Applied Microbiology and Biotechnology·Yanhong Li, Xi Chen
Jun 24, 2008·Glycoconjugate Journal·Kate KolesVladislav M Panin
Oct 3, 2003·Biochemical and Biophysical Research Communications·Norihiro KikuchiHisashi Narimatsu
Nov 3, 2001·Biochimica Et Biophysica Acta·V Vallejo-RuizA Harduin-Lepers
Sep 1, 2001·Biochimie·A Harduin-LepersP Delannoy
Nov 18, 1998·Current Opinion in Structural Biology·M MühlenhoffR Gerardy-Schahn
Oct 13, 2009·Nature Structural & Molecular Biology·Francesco V RaoNatalie C J Strynadka
Jun 18, 2010·Molecular BioSystems·Sara Fasmer HansenChristelle Breton
Jan 11, 2002·European Journal of Biochemistry·Martine MalissardThierry Hennet
Feb 27, 2003·European Journal of Biochemistry·Marie-Ange Krzewinski-RecchiPhilippe Delannoy
Dec 21, 2011·The Journal of Biological Chemistry·Joseph L Zapater, Karen J Colley
Dec 14, 2007·Glycobiology·Tatsuo MiyazakiMinoru Fukuda
Nov 26, 2010·Glycobiology·Magali AudryChristelle Breton
Nov 1, 2011·Journal of Biochemistry·Yuichi HiranoShuichi Tsuji
Mar 3, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Dong-Uk KimHyun-Soo Cho
May 8, 2008·Bioscience, Biotechnology, and Biochemistry·Shou Takashima
Dec 9, 2010·Marine Drugs·Takeshi Yamamoto
May 8, 2008·Neurochemical Research·Herbert HildebrandtRita Gerardy-Schahn
Jan 29, 2013·Neurochemical Research·Martina MühlenhoffHerbert Hildebrandt
Mar 20, 2008·Biochemical and Biophysical Research Communications·Yihua Gu, Robert K Yu
Jul 21, 2015·Nature Structural & Molecular Biology·Gesa VolkersNatalie C J Strynadka
Oct 6, 1999·Current Opinion in Structural Biology·C Breton, A Imberty
May 28, 2016·American Journal of Medical Genetics. Part a·Jin Sook LeeJong-Hee Chae
Dec 16, 2016·Histochemistry and Cell Biology·Gaurang P Bhide, Karen J Colley
Feb 3, 2006·Journal of Biochemistry·Shou TakashimaMasafumi Tsujimoto
Jan 3, 2001·The Journal of Biological Chemistry·H KawaiR L Proia
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