Abstract
ATP:cob(I)alamin adenosyltransferase (EutT) of Salmonella enterica was overproduced and enriched to approximately 70% homogeneity, and its basic kinetic parameters were determined. Abundant amounts of EutT protein were produced, but all of it remained insoluble. Soluble active EutT protein (approximately 70% homogeneous) was obtained after treatment with detergent. Under conditions in which cobalamin (Cbl) was saturating, Km(ATP) = 10 microm, kcat = 0.03 s(-1), and Vmax = 54.5 nm min(-1). Similarly, under conditions in which MgATP was saturating, Km(Cbl) = 4.1 microm, kcat = 0.06 s(-1), and Vmax = 105 nm min(-1). Unlike other ATP:co(I)rrinoid adenosyltransferases in the cell (i.e. CobA and PduO), EutT activity was > or =50-fold higher with ATP versus GTP, and EutT retained 80% of its activity with ADP substituted for ATP and was completely inactive with AMP as substrate, indicating that the enzyme requires the beta-phosphate group of the nucleotide substrate. The data suggest that the amino group of adenine might play a role in nucleotide recognition and/or binding. Unlike the housekeeping CobA enzyme, EutT was not inhibited by inorganic tripolyphosphate (PPPi). Results from 31P NMR spectroscopy studies identified PPi and Pi as...Continue Reading
References
Dec 1, 1976·Applied and Environmental Microbiology·W E Balch, R S Wolfe
Jan 1, 1990·Journal of Bacteriology·J C Escalante-SemerenaJ R Roth
Jul 1, 1968·Journal of Bacteriology·D BerkowitzB N Ames
Aug 15, 1970·Nature·U K Laemmli
Jul 1, 1968·The Journal of Pathology and Bacteriology·I F KeymerG Fretwell
May 1, 1984·Journal of Bacteriology·B A CastilhoM J Casadaban
Jul 1, 1995·Journal of Bacteriology·L M GuzmanJ Beckwith
Feb 1, 1995·Journal of Bacteriology·S Suh, J C Escalante-Semerena
Mar 1, 1995·Journal of Bacteriology·I StojiljkovicF Heffron
Jul 15, 1993·Gene·S J Suh, J C Escalante-Semerena
Mar 1, 1993·Journal of Bacteriology·V BianchiE Haggård-Ljungquist
Nov 14, 1997·Journal of Bacteriology·T A BobikJ R Roth
Aug 28, 1999·Journal of Bacteriology·E KofoidJ Roth
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Jan 10, 2001·Biochemistry·C B BauerI Rayment
Feb 13, 2001·Journal of Bacteriology·C L JohnsonT A Bobik
Jun 16, 2001·The Journal of Biological Chemistry·M V Fonseca, J C Escalante-Semerena
Sep 12, 2001·Biochemistry·Y KakutaK Fukuyama
Feb 15, 2002·Journal of Bacteriology·Gregory D HavemannThomas A Bobik
Jun 25, 2002·The Journal of Biological Chemistry·Maris V FonsecaJorge C Escalante-Semerena
Jan 7, 2003·The Journal of Biological Chemistry·Nicole A LealThomas A Bobik
Apr 11, 2003·The Journal of Biological Chemistry·Amanda A BrindleyMartin J Warren
Jan 20, 2004·Journal of Bacteriology·Giuseppe Bertani
Sep 1, 1951·Journal of Bacteriology·G BERTANI
Mar 27, 2004·The Journal of Biological Chemistry·Vivian SaridakisAled M Edwards
Aug 20, 2004·Journal of Bacteriology·Nicole R BuanJorge C Escalante-Semerena
Sep 7, 2004·The Journal of Biological Chemistry·Nicole A LealThomas A Bobik
Nov 2, 2004·Journal of Bacteriology·David E SheppardJohn R Roth
Nov 18, 2004·Journal of Bacteriology·Celeste L V JohnsonThomas A Bobik
Jun 16, 2005·Journal of the American Chemical Society·Troy A StichThomas C Brunold
Aug 6, 2005·Science·Cheryl A KerfeldTodd O Yeates
Nov 18, 2005·Journal of Bacteriology·Shaun R BrinsmadeJorge C Escalante-Semerena
Citations
Aug 3, 2010·Applied Microbiology and Biotechnology·Paola E Mera, Jorge C Escalante-Semerena
Sep 29, 1993·Biochimica Et Biophysica Acta·L M WorshamM L Ernst-Fonberg
Jan 4, 2013·The Journal of Physical Chemistry. a·Karina KornobisPawel M Kozlowski
Jul 30, 2011·Metallomics : Integrated Biometal Science·Lee Macomber, Robert P Hausinger
Nov 26, 2009·The Journal of Biological Chemistry·Paola E Mera, Jorge C Escalante-Semerena
Jul 27, 2010·Journal of Bacteriology·Shouqiang Cheng, Thomas A Bobik
Dec 7, 2007·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Pilar Ruiz-SánchezRoger Alberto
Aug 9, 2012·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Manoj KumarPawel M Kozlowski
Feb 18, 2016·Journal of the American Chemical Society·Ivan G PallaresThomas C Brunold
Nov 26, 2014·Biochemistry·Ivan G PallaresThomas C Brunold
Jan 4, 2017·Biochemistry·Ivan G PallaresThomas C Brunold
Dec 18, 2013·Journal of Bacteriology·Theodore C MooreJorge C Escalante-Semerena
Sep 4, 2014·Microbiology and Molecular Biology Reviews : MMBR·Chiranjit ChowdhuryThomas A Bobik
Nov 24, 2006·The Journal of Biological Chemistry·Martin St MauriceIvan Rayment
Oct 9, 2015·Molecular Microbiology·Theodore C Moore, Jorge C Escalante-Semerena
Jan 25, 2020·International Journal of Environmental Research and Public Health·Giuseppe GenchiAlessia Catalano
Jul 8, 2015·Molecular Microbiology·Thomas A BobikTodd O Yeates
Mar 2, 2021·Chemosphere·Amit KumarArti Bhatia
Sep 1, 2008·EcoSal Plus·Jorge C Escalante-Semerena, Martin J Warren
Oct 9, 2020·EcoSal Plus·Katie L StewartThomas A Bobik
Aug 4, 2018·Biochemistry·Flavia G Costa, Jorge C Escalante-Semerena
Mar 4, 2020·Biochemistry·Flavia G CostaJorge C Escalante-Semerena