What Kinesin Does at Roadblocks: the Coordination Mechanism for Molecular Walking

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2003 Dec 20

PMID 14685258

Citations 41

Authors

Isabelle M-T C Crevel

Miklos Nyitrai

Maria C Alonso

Stefan Weiss

Michael A Geeves

Robert A Cross

Affiliations

Soon will be listed here.

Abstract

Competing models for the coordination of processive stepping in kinesin can be tested by introducing a roadblock to prevent lead head attachment. We used T93N, an irreversibly binding mutant monomer, as a roadblock, and measured the rates of nucleotide-induced detachment of kinesin monomers or dimers with and without the T93N roadblock using microflash photolysis combined with stopped flow. Control nucleotide-induced monomer (rK340) unbinding was 73.6 s(-1) for ATP and 40.5 s(-1) for ADP. Control ADP-induced dimer (rK430) unbinding was 18.6 s(-1). Added 20 mM Pi slowed both monomer and dimer unbinding. With the roadblock in place, lead head attachment of dimers is prevented and ATP-induced trail head unbinding was then 42 s(-1). This is less than two-fold slower than the stepping rate of unimpeded rK430 dimers (50-70 s(-1)), indicating that during walking, lead head attachment induces at most only a slight (less than two-fold) acceleration of trail head detachment. As we discuss, this implies a coordination model having very fast (>2000 s(-1)) ATP-induced attachment of the lead head, followed by slower, strain-sensitive ADP release from the lead head.

Citing Articles

How Kinesin-1 Utilize the Energy of Nucleotide: The Conformational Changes and Mechanochemical Coupling in the Unidirectional Motion of Kinesin-1.

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These motors were made for walking.

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The ability of the kinesin-2 heterodimer KIF3AC to navigate microtubule networks is provided by the KIF3A motor domain.

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Kinesin-2 motors: Kinetics and biophysics.

Gilbert S, Guzik-Lendrum S, Rayment I J Biol Chem. 2018; 293(12):4510-4518.

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