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Frustrated binding of biopolymer crosslinkers

Yuval Mulla, Harmen Wierenga, Celine Alkemade, Pieter Rein ten Wolde, Gijsje H. Koenderink
doi: https://doi.org/10.1101/483669
Yuval Mulla
1Living Matter Department, AMOLF, Science Park 104, 1098 XG Amsterdam
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Harmen Wierenga
1Living Matter Department, AMOLF, Science Park 104, 1098 XG Amsterdam
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Celine Alkemade
1Living Matter Department, AMOLF, Science Park 104, 1098 XG Amsterdam
2Department of Bionanoscience, Faculty of Applied Sciences, Delft University of Technology, Delft, Netherlands
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Pieter Rein ten Wolde
1Living Matter Department, AMOLF, Science Park 104, 1098 XG Amsterdam
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Gijsje H. Koenderink
1Living Matter Department, AMOLF, Science Park 104, 1098 XG Amsterdam
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  • For correspondence: g.koenderink@amolf.nl
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Abstract

Transiently crosslinked actin filament networks allow cells to combine elastic rigidity with the ability to deform viscoelastically. Theoretical models of semiflexible polymer networks predict that the crosslinker unbinding rate governs the timescale beyond which viscoelastic flow occurs. However a direct comparison between network and crosslinker dynamics is lacking. Here we measure the network’s stress relaxation timescale using rheology and the lifetime of bound crosslinkers using fluorescence recovery after photobleaching. lntruigingly, we observe that the crosslinker unbinding rate measured by FRAP is more than an order of magnitude slower than the rate measured by rheology. We rationalize this difference with a three-state model where crosslinkers are bound to either 0, 1 or 2 filaments, which allows us to extract crosslinker transition rates that are otherwise difficult to access. We find that the unbinding rate of singly bound crosslinkers is nearly two orders of magnitude slower than for double bound ones. We attribute the increased unbinding rate of doubly bound crosslinkers to the high stiffness of biopolymers, which frustrates crosslinker binding.

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Posted November 30, 2018.
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Frustrated binding of biopolymer crosslinkers
Yuval Mulla, Harmen Wierenga, Celine Alkemade, Pieter Rein ten Wolde, Gijsje H. Koenderink
bioRxiv 483669; doi: https://doi.org/10.1101/483669
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Frustrated binding of biopolymer crosslinkers
Yuval Mulla, Harmen Wierenga, Celine Alkemade, Pieter Rein ten Wolde, Gijsje H. Koenderink
bioRxiv 483669; doi: https://doi.org/10.1101/483669

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