Spectrophotometric Assays to Quantify the Activity of T4SS ATPases

Elena Cabezón; Ignacio Arechaga

doi:10.1007/978-1-4939-9877-7_10

Spectrophotometric Assays to Quantify the Activity of T4SS ATPases

Methods Mol Biol. 2020:2075:135-143. doi: 10.1007/978-1-4939-9877-7_10.

Authors

Elena Cabezón¹, Ignacio Arechaga²

Affiliations

¹ Departamento de Biología Molecular, Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC), Universidad de Cantabria-CSIC, Santander, Spain. cabezone@unican.es.
² Departamento de Biología Molecular, Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC), Universidad de Cantabria-CSIC, Santander, Spain.

PMID: 31584160
DOI: 10.1007/978-1-4939-9877-7_10

Abstract

Biogenesis of T4SS apparatus and substrate transport require energy. Conjugative T4SS have three ATPases that enable DNA processing and transport of the nucleoprotein complex to the recipient cell. In the conjugative plasmid R388, these ATPases are named TrwB, TrwK, and TrwD. Here, three different spectrophotometric assays to measure the enzymatic properties of these ATPases are described. The choice of the assay will depend on the specific requirements of each enzyme.

Keywords: ATPase activity; Coupled assay; Molecular motor; Spectrophotometric assay.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine Triphosphatases / metabolism*
Bacterial Proteins / metabolism
Enzyme Activation
Hydrolysis
Spectrophotometry* / methods
Type V Secretion Systems / metabolism*

Substances

Bacterial Proteins
Type V Secretion Systems
Adenosine Triphosphatases