Abstract
We studied the functional role of the Escherichia coli ADPglucose pyrophosphorylase's N-terminus in allosteric regulation, and the particular effects caused by its length. Small truncated mutants were designed, and those lacking up to 15-residues were active and highly purified for further kinetic analyses. Ndelta3 and Ndelta7 did not change the kinetic parameters with respect to the wild-type. Ndelta11 and Ndelta15 enzymes were insensitive to allosteric regulation and highly active in the absence of the activator. Co-expression of two polypeptides corresponding to the N- and C-termini generated an enzyme with activation properties lower than those of the wild-type [C.M. Bejar, M.A. Ballicora, D.F. Gómez Casati, A.A. Iglesias, J. Preiss, The ADPglucose pyrophosphorylase from Escherichia coli comprises two tightly bound distinct domains, FEBS Lett. 573 (2004) 99-104]. Here, we characterized a Ndelta15 co-expression mutant, in which the allosteric regulation was restored to wild-type levels. Unusual allosteric effects caused by either an N-terminal truncation or co-expression of individual domains may respond to structural changes favoring an up-regulated or a down-regulated conformation rather than specific activator or inhibito...Continue Reading
References
Jul 1, 1990·Archives of Biochemistry and Biophysics·A Gardiol, J Preiss
Oct 1, 1985·Analytical Biochemistry·P K SmithD C Klenk
Jun 1, 1966·Biochemistry·J PreissN Gentner
Aug 15, 1970·Nature·U K Laemmli
Sep 29, 1998·Archives of Biochemistry and Biophysics·M X Wu, J Preiss
May 8, 2001·Archives of Biochemistry and Biophysics·M X Wu, J Preiss
Aug 22, 2001·Biochemistry·D F Gómez-CasatiC R Meyer
Sep 22, 2001·The Journal of Biological Chemistry·J B FrueaufJ Preiss
Jul 24, 2002·Biochemistry·Miguel A BallicoraJack Preiss
Jun 10, 2003·Microbiology and Molecular Biology Reviews : MMBR·Miguel A BallicoraJack Preiss
Dec 5, 2003·Analytical Biochemistry·Alejandra YepJack Preiss
Aug 26, 2004·FEBS Letters·Clarisa M BejarJack Preiss
Feb 5, 2005·The EMBO Journal·Xiangshu JinJames H Geiger
Oct 18, 2005·Photosynthesis Research·Miguel A BallicoraJack Preiss
Sep 1, 1987·Plant Physiology·M K MorellJ Preiss
Oct 9, 1992·Science·D M StarkG M Kishore
Citations
Jul 5, 2013·PloS One·Carlos M FigueroaAlberto A Iglesias
Dec 1, 2015·Frontiers in Microbiology·Ana C EbrechtMiguel A Ballicora
Jul 12, 2011·Biochimie·Carlos M FigueroaMiguel A Ballicora
Oct 26, 2016·Protein Science : a Publication of the Protein Society·Eszter NémethBéla Gyurcsik
May 8, 2013·International Journal of Molecular Sciences·Matías D Asención DiezAlberto A Iglesias
Jan 27, 2015·Protein Science : a Publication of the Protein Society·Benjamin L HillMiguel A Ballicora
Nov 3, 2006·The Journal of Biological Chemistry·Clarisa Maria BejarJack Preiss
May 1, 2014·EcoSal Plus·Jack Preiss
Aug 1, 2009·EcoSal Plus·Jack Preiss
Jun 16, 2021·Journal of Zhejiang University. Science. B·Yun WuYiping Xia