Mechanical cues often influence the factors affecting the transition states of catalytic reactions and alter the activation pathway. However, tracking the real-time dynamics of such activation pathways is limited. Using single-molecule trapping of reaction intermediates, we developed a method that enabled us to perform one reaction at one site and simultaneously study the real-time dynamics of the catalytic pathway. Using this, we showed single-molecule calligraphy at nanometer resolution and deciphered the mechanism of the sortase A enzymatic reaction that, counter-intuitively, accelerates bacterial adhesion under shear tension. Our method captured a force-induced dissociation of the enzyme–substrate bond that accelerates the forward reaction 100×, proposing a new mechano-activated catalytic pathway. In corroboration, our molecular dynamics simulations in the presence of force identified a force-induced conformational switch in the enzyme that accelerates proton transfer between CYS184 (acceptor) and HIS120 (donor) catalytic dyads by reducing the inter-residue distances. Overall, the present study opens up the possibility of studying the influence of factors affecting transition states in real time and paves the way for the rational design of enzymes with enhanced efficiency.
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Cover Image
A schematic representation of the steps in involved in single-molecule calligraphy using the TERI (trapping the enzymatic reaction intermediate) technique used by Hazra et al. to track force-activated reaction pathways in real time. In this issue, these authors report on the mechanism of the sortase A enzymatic reaction that, counter-intuitively, accelerates bacterial adhesion under shear tension. For details, see pages 2611–2620.
Research Article|
August 29 2018
Force-activated catalytic pathway accelerates bacterial adhesion against flow
Jagadish P. Hazra
;
Jagadish P. Hazra
1Department of Chemical Sciences, Indian Institute of Science Education and Research Mohali, Punjab 140306, India
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Nisha Arora
;
Nisha Arora
*
1Department of Chemical Sciences, Indian Institute of Science Education and Research Mohali, Punjab 140306, India
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Amin Sagar
;
Amin Sagar
*
1Department of Chemical Sciences, Indian Institute of Science Education and Research Mohali, Punjab 140306, India
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Shwetha Srinivasan
;
Shwetha Srinivasan
1Department of Chemical Sciences, Indian Institute of Science Education and Research Mohali, Punjab 140306, India
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Abhishek Chaudhuri
;
Abhishek Chaudhuri
2Department of Physical Sciences, Indian Institute of Science Education and Research Mohali, Punjab 140306, India
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Sabyasachi Rakshit
1Department of Chemical Sciences, Indian Institute of Science Education and Research Mohali, Punjab 140306, India
3Centre for Protein Science Design and Engineering, Indian Institute of Science Education and Research Mohali, Punjab 140306, India
Correspondence: Sabyasachi Rakshit (srakshit@iisermohali.ac.in)
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Biochem J (2018) 475 (16): 2611–2620.
Article history
Received:
May 13 2018
Revision Received:
June 28 2018
Accepted:
July 02 2018
Accepted Manuscript online:
July 02 2018
Citation
Jagadish P. Hazra, Nisha Arora, Amin Sagar, Shwetha Srinivasan, Abhishek Chaudhuri, Sabyasachi Rakshit; Force-activated catalytic pathway accelerates bacterial adhesion against flow. Biochem J 31 August 2018; 475 (16): 2611–2620. doi: https://doi.org/10.1042/BCJ20180358
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