Single alkaline phosphatase molecule assay by capillary electrophoresis laser-induced fluorescence detection

Douglas B. Craig, Edgar A. Arriaga, Jerome C.Y. Wong, Hui Lu, Norman J. Dovichi

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Single molecules of alkaline phosphatase were assayed using capillary electrophoresis laser-induced fluorescence detection. Multiple incubations of individual molecules were performed. Varying the temperature in multiple incubation assay allowed for the determination of the activation energy of catalysis at the single molecule level. Molecules are heterogeneous with respect to both activity and activation energy of catalysis. Partial thermal denaturation of alkaline phosphatase results from the total denaturation of a fraction of the molecules with surviving molecules unaffected rather than a partial decrease in the activity of all the molecules.

Original languageEnglish (US)
Pages (from-to)121-131
Number of pages11
JournalTechniques in Protein Chemistry
Volume8
Issue numberC
DOIs
StatePublished - Dec 1 1997

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