Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputs

Pushpanjali Soppina, Nishaben Patel, Dipeshwari J. Shewale, Ashim Rai, Sivaraj Sivaramakrishnan, Pradeep K. Naik, Virupakshi Soppina

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


BACKGROUND: Kinesin-3 family motors drive diverse cellular processes and have significant clinical importance. The ATPase cycle is integral to the processive motility of kinesin motors to drive long-distance intracellular transport. Our previous work has demonstrated that kinesin-3 motors are fast and superprocessive with high microtubule affinity. However, chemomechanics of these motors remain poorly understood.

RESULTS: We purified kinesin-3 motors using the Sf9-baculovirus expression system and demonstrated that their motility properties are on par with the motors expressed in mammalian cells. Using biochemical analysis, we show for the first time that kinesin-3 motors exhibited high ATP turnover rates, which is 1.3- to threefold higher compared to the well-studied kinesin-1 motor. Remarkably, these ATPase rates correlate to their stepping rate, suggesting a tight coupling between chemical and mechanical cycles. Intriguingly, kinesin-3 velocities (KIF1A > KIF13A > KIF13B > KIF16B) show an inverse correlation with their microtubule-binding affinities (KIF1A < KIF13A < KIF13B < KIF16B). We demonstrate that this differential microtubule-binding affinity is largely contributed by the positively charged residues in loop8 of the kinesin-3 motor domain. Furthermore, microtubule gliding and cellular expression studies displayed significant microtubule bending that is influenced by the positively charged insert in the motor domain, K-loop, a hallmark of kinesin-3 family.

CONCLUSIONS: Together, we propose that a fine balance between the rate of ATP hydrolysis and microtubule affinity endows kinesin-3 motors with distinct mechanical outputs. The K-loop, a positively charged insert in the loop12 of the kinesin-3 motor domain promotes microtubule bending, an interesting phenomenon often observed in cells, which requires further investigation to understand its cellular and physiological significance.

Original languageEnglish (US)
Article number177
JournalBMC Biology
Issue number1
StatePublished - Dec 2022

Bibliographical note

Funding Information:
V.S. acknowledges funding through DBT (Grant No.: BT/PR15214/BRB/10/1449/2015 and BT/RLF/Re-entry/45/2015) and DST-SERB (Grant No.: ECR/2016/000913). P.K.N acknowledges ICMR for funding (Grant No. 5/13/13/2019/NCD-III). P.S. acknowledges funding from DST (Grant No.: SR/WOS-A/LS-73/2017). N.M.P and D.J.S acknowledges fellowship from IIT Gandhinagar.

Publisher Copyright:
© 2022, The Author(s).


  • ATPases
  • Baculovirus
  • Chemomechanical
  • Kinesin-3
  • Microtubule bending
  • Superprocessive
  • Adenosine Triphosphate/metabolism
  • Kinesins/genetics
  • Mammals
  • Adenosine Triphosphatases/analysis
  • Microtubules/metabolism
  • Animals
  • Protein Binding

PubMed: MeSH publication types

  • Research Support, Non-U.S. Gov't
  • Journal Article


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