Bottom-up single-molecule strategy for understanding subunit function of tetrameric β-galactosidase.

Abstract:

:In this paper, we report an example of the engineered expression of tetrameric β-galactosidase (β-gal) containing varying numbers of active monomers. Specifically, by combining wild-type and single-nucleotide polymorphism plasmids at varying ratios, tetrameric β-gal was expressed in vitro with one to four active monomers. The kinetics of individual enzyme molecules revealed four distinct populations, corresponding to the number of active monomers in the enzyme. Using single-molecule-level enzyme kinetics, we were able to measure an accurate in vitro mistranslation frequency (5.8 × 10-4 per base). In addition, we studied the kinetics of the mistranslated β-gal at the single-molecule level.

authors

Li X,Jiang Y,Chong S,Walt DR

doi

10.1073/pnas.1805690115

subject

Has Abstract

pub_date

2018-08-14 00:00:00

pages

8346-8351

issue

33

eissn

0027-8424

issn

1091-6490

pii

1805690115

journal_volume

115

pub_type

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