Spectrophotometric Assays to Quantify the Activity of T4SS ATPases

  • Elena CabezónEmail author
  • Ignacio Arechaga
Part of the Methods in Molecular Biology book series (MIMB, volume 2075)


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.

Key words

ATPase activity Molecular motor Coupled assay Spectrophotometric assay 



This work was supported by the Spanish Ministerio de Economía y Competitividad (MINECO) grants BFU2016-78521-R (to E.C. and I.A.) and BFU2014-61823-EXP (to E.C.).


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Departamento de Biología Molecular, Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC)Universidad de Cantabria-CSICSantanderSpain

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