Abstract
Crystal structures of adenylosuccinate synthetase from Escherichia coli complexed with Mg2+, 6-thiophosphoryl-IMP, GDP, and hadacidin at 298 and 100 K have been refined to R-factors of 0.171 and 0.206 against data to 2.8 and 2.5 A resolution, respectively. Interactions of GDP, Mg2+ and hadacidin are similar to those observed for the same ligands in the complex of IMP, GDP, NO3-, Mg2+ and hadacidin (Poland, B. W., Fromm, H. J. & Honzatko, R. B. (1996). J. Mol. Biol. 264, 1013-1027). Although crystals were grown from solutions containing 6-mercapto-IMP and GTP, the electron density at the active site is consistent with 6-thiophosphoryl-IMP and GDP. Asp-13 and Gln-224 probably work in concert to stabilize the 6-thioanion of 6-mercapto-IMP, which in turn is the nucleophile in the displacement of GDP from the gamma-phosphate of GTP. Once formed, 6-thiophosphoryl-IMP is stable in the active site of the enzyme under the conditions of the structural investigation. The direct observation of 6-thiophosphoryl-IMP in the active site is consistent with the putative generation of 6-phosphoryl-IMP along the reaction pathway of the synthetase.
References
Nov 1, 1977·Archives of Biochemistry and Biophysics·G D Markham, G H Reed
Jun 23, 1976·Biochimica Et Biophysica Acta·A W Clark, F B Rudolph
Jul 1, 1987·Archives of Biochemistry and Biophysics·M B BassM M Stayton
Nov 18, 1986·Biochemistry·B F CooperF B Rudolph
Jan 1, 1985·Methods in Enzymology·A J HowardN H Xuong
Jan 1, 1983·Current Topics in Cellular Regulation·M M StaytonH J Fromm
Aug 1, 1983·Archives of Biochemistry and Biophysics·D J PorterH J Bright
Jan 15, 1982·Biochemical Pharmacology·T SpectorJ J Marr
Dec 1, 1995·Journal of Molecular Biology·M M SilvaR B Honzatko
Jun 28, 1996·The Journal of Biological Chemistry·B W PolandR B Honzatko
Dec 10, 1996·Biochemistry·B W PolandR B Honzatko
Dec 20, 1996·Journal of Molecular Biology·B W PolandR B Honzatko
May 2, 1997·The Journal of Biological Chemistry·C KangH J Fromm
Aug 1, 1958·Biochimica Et Biophysica Acta·H J FROMM
Jul 1, 1963·Journal of Molecular Biology·G N RAMACHANDRANV SASISEKHARAN
Mar 1, 1962·Biochemistry·E A KACZKAK FOLKERS
Citations
Nov 24, 1999·Archives of Biochemistry and Biophysics·P LeeR F Colman
Dec 6, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Xiaoying WangYong Xie
Oct 27, 2007·Biochimie·Stéphane Descorps-DeclèreBernard Labedan
Apr 7, 2016·Biochemistry·Vishakha KarnawatMrinalini Puranik
Jul 15, 2015·Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy·Vishakha Karnawat, Mrinalini Puranik
Jun 20, 1998·The Journal of Biological Chemistry·W WangH J Fromm
May 11, 2002·The Journal of Biological Chemistry·Cristina V IancuRichard B Honzatko
Nov 2, 2013·Chemical Communications : Chem Comm·Natsuhisa OkaKaori Ando
Sep 19, 2001·The Journal of Biological Chemistry·C V IancuR B Honzatko
Jan 10, 2002·The Journal of Biological Chemistry·Andrea GorrellHerbert J Fromm
Aug 12, 2020·ChemMedChem·Nidhi Tibrewal, Gregory I Elliott
Jul 4, 1997·The Journal of Biological Chemistry·W WangH J Fromm
Jul 6, 2019·European Biophysics Journal : EBJ·Tadeo E SaldañoSebastian Fernandez-Alberti
Jun 11, 1999·The Journal of Biological Chemistry·Z HouR B Honzatko
Dec 14, 2001·The Journal of Biological Chemistry·Zhenglin HouRichard B Honzatko
Aug 21, 2002·The Journal of Biological Chemistry·Cristina V IancuRichard B Honzatko
May 12, 2006·The Journal of Biological Chemistry·Nathaniel D GinderRichard B Honzatko
Jan 12, 2005·Biochemistry·Scott W NelsonHerbert J Fromm
May 29, 1999·Biochemistry·J Y ChoeR B Honzatko
Oct 13, 2007·Biochemistry·Sonali Mehrotra, Hemalatha Balaram
Sep 25, 1999·Archives of Biochemistry and Biophysics·R B Honzatko, H J Fromm
Sep 1, 2008·EcoSal Plus·Kaj Frank JensenMartin WillemoËs
Jun 24, 1998·Biochemistry·J B ThodenH M Holden
May 8, 1999·Biochemistry·P LeeR F Colman
Sep 20, 2006·Biochemistry·Cristina V IancuRichard B Honzatko