Adhesion force and attachment lifetime of the KIF16B-PX domain interaction with lipid membranes

Mol Biol Cell. 2017 Nov 7;28(23):3315-3322. doi: 10.1091/mbc.E17-05-0324. Epub 2017 Sep 20.

Abstract

KIF16B is a highly processive kinesin-3 family member that participates in the trafficking and tubulation of early endosomes along microtubules. KIF16B attaches to lipid cargoes via a PX motif at its C-terminus, which has nanomolar affinity for bilayers containing phosphatidylinositol-3-phosphate (PI[3]P). As the PX domain has been proposed to be a primary mechanical anchor for the KIF16B-cargo attachment, we measured the adhesion forces and detachment kinetics of the PX domain as it interacts with membranes containing 2% PI(3)P and 98% phosphatidylcholine. Using optical tweezers, we found that the adhesion strength of a single PX domain ranged between 19 and 54 pN at loading rates between 80 and 1500 pN/s. These forces are substantially larger than the interaction of the adhesion of a pleckstrin homology domain with phosphatidylinositol 4,5-bisphosphate. This increased adhesion is the result of the membrane insertion of hydrophobic residues adjacent to the PI(3)P binding site, in addition to electrostatic interactions with PI(3)P. Attachment lifetimes under load decrease monotonically with force, indicating slip-bond behavior. However, the lifetime of membrane attachment under load appears to be well matched to the duration of processive motility of the KIF16B motor, indicating the PX domain is a suitable mechanical anchor for intracellular transport.

MeSH terms

  • Binding Sites / physiology
  • Biological Transport
  • Cell Adhesion / physiology
  • Cell Membrane / metabolism
  • Endosomes / metabolism
  • Kinesin / metabolism*
  • Kinesin / physiology*
  • Kinetics
  • Membrane Lipids / metabolism
  • Microtubules / metabolism
  • Models, Molecular
  • Phosphatidylinositol Phosphates / metabolism
  • Phosphatidylinositol Phosphates / physiology
  • Phosphatidylinositols / metabolism
  • Protein Binding / physiology
  • Protein Domains / physiology
  • Protein Transport

Substances

  • KIF16B protein, human
  • Membrane Lipids
  • Phosphatidylinositol Phosphates
  • Phosphatidylinositols
  • phosphatidylinositol 3-phosphate
  • Kinesin