Abstract
Biophysical studies of the mechanochemical cycle of kinesin motors are essential for understanding the mechanism of energy conversion. Here, we report a systematic study of the impact of temperature on velocity and run length of homodimeric Drosophila kinesin-1, homodimeric C. elegans OSM-3 and heterodimeric C. elegans kinesin-II motor proteins using in vitro single-molecule motility assays. Under saturated ATP conditions, kinesin-1 and OSM-3 are fast and processive motors compared to kinesin-II. From in vitro motility assays employing single-molecule fluorescence microscopy, we extracted single-motor velocities and run lengths in a temperature range from 15 °C to 35 °C. Both parameters showed a non-Arrhenius temperature dependence for all three motors, which could be quantitatively modeled using a simplified, two-state kinetic model of the mechanochemistry of the three motors, providing new insights in the temperature dependence of their mechanochemistry.
| Original language | English |
|---|---|
| Pages (from-to) | 812-818 |
| Number of pages | 7 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 529 |
| Issue number | 3 |
| Early online date | 30 Jun 2020 |
| DOIs | |
| Publication status | Published - 27 Aug 2020 |
Funding
This work was financially supported by the Netherlands Foundation for Scientific Research (NWO) via the grant “Barriers in the Brain” ( BIB, FOM-137 ) from the Foundation for Fundamental Research on Matter ( FOM ).
| Funders | Funder number |
|---|---|
| Netherlands Foundation for Scientific Research | |
| Stichting voor Fundamenteel Onderzoek der Materie | |
| Nederlandse Organisatie voor Wetenschappelijk Onderzoek | FOM-137 |
| Foundation for Fundamental Research on Matter |
Keywords
- ATP hydrolysis cycle
- Caenorhabditis elegans
- Kinesins
- Microtubules
- Motor proteins
- Single molecule analysis
- Single-molecule fluorescence microscopy
- Temperature
- TIRF microscopy