The Thumb Domain is Not Essential for the Catalytic Action of HoLaMa DNA Polymerase

Overview

Journal Protein J

Publisher Springer

Specialty Biochemistry

Date 2017 Sep 22

PMID 28932939

Citations 1

Authors

Angela Gala Morena Gatius

Fabrizio Dal Piaz

Alejandro Hochkoeppler

Affiliations

Soon will be listed here.

Abstract

A structural and kinetic characterization of a fragment of the HoLaMa DNA polymerase is presented here. In particular, a truncated form of HoLaMa, devoid of a consistent portion of the thumb domain, was isolated and purified. This HoLaMa fragment, denoted as ΔNter-HoLaMa, is surprisingly competent in catalyzing DNA extension, albeit featuring a k one order of magnitude lower than the corresponding kinetic constant of its full-length counterpart. The conformational rearrangements, if any, of enzyme tryptophanes triggered by DNA binding or extension were assayed under pre-steady-state conditions. The fluorescence of HoLaMa tryptophanes was found to significantly change upon DNA binding and extension. On the contrary, no fluorescence changes of ΔNter-HoLaMa tryptophanes were detected under the same conditions, suggesting that major conformational transitions are not required for DNA binding or extension by this truncated DNA polymerase.

Citing Articles

Structural and catalytic insights into HoLaMa, a derivative of Klenow DNA polymerase lacking the proofreading domain.

Kovermann M, Stefan A, Castaldo A, Caramia S, Hochkoeppler A PLoS One. 2019; 14(4):e0215411.

PMID: 30970012 PMC: 6457538. DOI: 10.1371/journal.pone.0215411.

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