期刊
METHODS
卷 204, 期 -, 页码 348-360出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2021.12.002
关键词
Magnetic tweezers; DNA helicase; Optical trap; SMARCAL1; Pif1; RecG
资金
- NIH [GM100156]
This review discusses important aspects of studying DNA helicases, including the use of various assays, the contribution of single-molecule studies, and the correlation with bulk-phase studies.
There are multiple assays available that can provide insight into the biochemical mechanism of DNA helicases. For the first 22 years since their discovery, bulk-phase assays were used. These include gel-based, spectrophotometric, and spectrofluorometric assays that revealed many facets of these enzymes. From 2001, singlemolecule studies have contributed additional insight into these DNA nanomachines to reveal details on energy coupling, step size, processivity as well as unique aspects of individual enzyme behavior that were masked in the averaging inherent in ensemble studies. In this review, important aspects of the study of helicases are discussed including beginning with active, nuclease-free enzyme, followed by several bulk-phase approaches that have been developed and still find widespread use today. Finally, two single-molecule approaches are discussed, and the resulting findings are related to the results obtained in bulk-phase studies.
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