181
Views
3
CrossRef citations to date
0
Altmetric
BIOANALYTICAL

Single Molecule Assay of Escherichia coli β-Galactosidase Using Two Competing Substrates Simultaneously, DDAO-β-D-Galactoside and Resorufin-β-D-Galactoside

, , &
Pages 1835-1841 | Received 05 May 2010, Accepted 28 Jul 2010, Published online: 20 Jul 2011

REFERENCES

  • Chen , D. , H. R. Harke , and N. J. Dovichi . 1992 . Two-label peak-height encoded DNA sequencing by capillary gel electrophoresis: Three examples . Nucleic Acids Res. 20 : 4873 – 4880 .
  • Craig , D. B. , E. A. Arriaga , J. C. Y. Wong , H. Lu , and N. J. Dovichi . 1996 . Studies on single alkaline phosphatase molecules: Reaction rate and activation energy of a reaction catalyzed by a single molecule and the effect of thermal denaturation-The death of an enzyme . J. Am. Chem. Soc. 118 : 5245 – 5253 .
  • Craig , D. B. , and N. J. Dovichi . 1998 . Escherichia coli β-galactosidase is heterogeneous with respect to the activity of individual molecules . Can. J. Chem. 76 : 623 – 626 .
  • Craig , D. B. , T. Hall , and D. M. Goltz . 2000 . E. coli β-galactosidase is heterogeneous with respect to a requirement for magnesium . BioMetals 13 : 223 – 229 .
  • Craig , D. B. , A. M. Haslam , J. M. L. Coombs , and E. R. Nichols . 2010 . Kinetic studies of unmodified individual Escherichia coli β-galactosidase molecules in free solution . Biochem. Cell Biol. 88 : 451 – 458 .
  • Craig , D. B. , and E. R. Nichols . 2008. Continuous flow assay for the simultaneous measurement of the electrophoretic mobility, catalytic activity and its variation over time of individual molecules of Escherichia coli β-galactosidase. Electrophoresis 29: 4298–4303.
  • Edman , L , Z. Foldes-Papp , S. Wennmalm , and R. Rigler . 1999 . The fluctuating enzyme: A single molecule approach . Chem. Phys. 247 : 11 – 22 .
  • English , B. P. , W. Min , A. M. van Oijen , K. T. Lee , G. Luo , H. Sun , B. J. Cherayil , S. C. Kou , and X. S. Xie . 2006 . Ever-fluctuating single enzyme molecules: Michaelis-Menten equation revisited . Nature Chem. Biol. 2 : 87 – 94 .
  • Lu , H. P. , L. Sun , and X. S. Xie . 1998 . Single-molecule Enzymatic Dynamics . Science 282 : 1877 – 1882 .
  • Nichols , E. R. , and D. B. Craig . 2008 . Measurement of the differences in electrophoretic mobilities of individual molecules of E. coli β-galactosidase provides insight into structural differences which underlie enzyme microheterogeneity . Electrophoresis 2 9: 4257 – 4269 .
  • Nichols , E. R. , J. M. A. Gavina , R. G. McLeod , and D. B. Craig . 2007 . Single molecule assays of β-galactosidase from two wild-type strains of E. coli: Effects of protease inhibitors on microheterogeneity and different relative activities with differing substrates . Protein J. 26 : 95 – 105 .
  • Nichols , E. R. , E. Shadabi , and D. B. Craig . 2009 . Effect of translation error rate on enzymatic heterogeneity as assessed by variation in single molecule elecrrophoretic mobility and catalytic activity . Biochem. Cell Biol. 87 : 517 – 529 .
  • Rissin , D. M. , H. H. Gorris , and D. R. Walt . 2008 . Distinct and long lived activity states of single enzyme molecules . J. Am. Chem. Soc. 130 : 5349 – 5353 .
  • Shoemaker , G. K. , D. H. Juers , J. M. L. Coombs , B. W. Matthews , and D. B. Craig . 2003 . Crystallization of β-galactosidase does not reduce the range of activity of individual molecules . Biochemistry 42 : 1707 – 1710 .
  • Xue , Q. , and E. S. Yeung . 1995 . Differences in the chemical reactivity of individual molecules of an enzyme . Nature 373 : 681 – 683 .
  • Zhao , J. -Y. , D. -Y. Chen , and N. J. Dovichi . 1992 . Low-cost laser-induced fluorescence detector for micellar capillary zone electrophoresis: Detection at the zeptomolar level of tetramethylrhodamine thiocarbamyl amino acid derivatives . J. Chromatogr. 608 : 117 – 120 .

Reprints and Corporate Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

To request a reprint or corporate permissions for this article, please click on the relevant link below:

Academic Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

Obtain permissions instantly via Rightslink by clicking on the button below:

If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.