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Using molecular motors to extract lipid tubules

A Simple Experimental Model to Investigate Force Range for Membrane Nanotube Formation

Chai Lor1, Joseph D. Lopes2, Michelle K. Mattson-Hoss3, Jing Xu2* and Linda S. Hirst2*
  • 1Biological Engineering and Small-scale Technologies Graduate Program, School of Engineering, University of California Merced, Merced, CA, USA
  • 2Physics Department, School of Natural Science, University of California Merced, Merced, CA, USA
  • 3Developmental and Cell Biology, School of Biological Sciences, University of California Irvine, Irvine, CA, USA

fmats-03-00006-g001The presence of membrane tubules in living cells is essential to many biological processes. In cells, one mechanism to form nanosized lipid tubules is via molecular motor induced bilayer extraction. In this paper, we describe a simple experimental model to investigate the forces required for lipid tube formation using kinesin motors anchored to 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) vesicles. Previous related studies have used molecular motors actively pulling on the membrane to extract a nanotube. Here, we invert the system geometry; molecular motors are used as static anchors linking DOPC vesicles to a two-dimensional microtubule network and an external flow is introduced to generate nanotubes facilitated by the drag force. We found that a drag force of ≈7 pN was sufficient for tubule extraction for vesicles ranging from 1 to 2 μm in radius. By our method, we found that the force generated by a single molecular motor was sufficient for membrane tubule extraction from a spherical lipid vesicle.

Congratulations Dr Lor!

Dr Chai Lor successfully defended his PhD thesis in the BEST (Bioengineering and small scale technologies) program on October 26th. Chai was a bioengineering undergraduate at UC Merced and a member of the first graduating class.

“Phase Behavior and Nanotube Formation in Lipid Membranes”

Biological cells are protected by a complex barrier called the lipid membrane. The lipid membrane is a soft material structure consisting of many lipid molecules held together by hydrophobic forces in an aqueous solution. Two simple experimental models were employed to investigate the role of specific lipid molecules in biological membranes. The first model investigated the phase behavior of the binary lipid mixture, 1-dipalmitoyl-2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine (DHA-PE) and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), using small-angle x-ray scattering (SAXS) and wide-angle x-ray scattering (WAXS). Our results shows that DHA-PE induces phase separation into a DHA rich liquid crystalline (Lα) phase and a DHA poor gel (Lβ’) phase at overall DHA-PE concentrations as low as 0.1mol%. In addition, we find that the structure of the Lβ’ phase, from which the DHA-PE molecules are largely excluded, is modified in the phase-separated state at low DHA-PE concentrations, with a decrease in bilayer thickness of 1.34nm for 0.1mol% at room temperature compared to pure DPPC bilayers. The second model investigated the formation of lipid nanotubes using an anchor system consisting of lipids, kinesin molecular motors, and microtubules in a flow cell. Lipid tubulation was conducted on two different membranes, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and DPPC. Lipid nanotubes were pulled from anchored giant unilamellar vesicles (GUVs) by drag force generated from the flow inside the channel. The results showed that DPPC membranes cannot generate lipid nanotubes while DOPC can, which was expected. We find that a drag force of approximately ≈7.9 pN is sufficient for tubule extraction and that it only requires 1-2 kinesin motor proteins for anchoring the GUV.

“Low concentrations of docosahexanoic acid significantly modify membrane structure and phase behavior”
C. Lor and L.S. Hirst, MEMBRANES, 5(4), 857-874 Link (2015)

 

Kyle Kabasaras, CAMP summer student, presents research at UC Merced Symposium

This summer Kyle Kabasaras presented his original research project at the UC Merced undergrad research symposium.

Investigating quantum dot assembly in a cholesteric liquid crystal

An ongoing goal in condensed matter physics is directly controlling the self-assembly of quantum dots (QDs) into specific structures while maintaining their original electronic and optical properties. One method of controlling the self-assembly of QDs is to disperse them within a liquid crystal (LC) medium and apply a variety of thermal stimulations. Recently, our lab developed a method of creating spherical, vesicle-shaped QDs within a nematic LCVesicle formation depends on the QD concentration in the LC as well as the LC’s intermolecular dispersion forces and thermal properties. In this project, we investigate the dispersion of CdSe/ZnS (core/shell) QDs in a cholesteric LC (CLC) medium and predict the QD aggregations to cluster near the LC defects. By varying parameters such as QD concentration and temperature, we exploit the CLC’s sensitive optical and thermal propertiesTo observe these effects, we apply spectrophotometry, polarized optical microscopy, and fluorescence microscopy. These techniques highlight the aggregation of QDs within the host CLC and identify how LC phase transitions determine where QDaggregates form. This work illustrates the possibility of new LC-based QD devices, and we will continue by exploring the lasing potential of our sample.

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CAMP is a National Science Foundation funded project to promote undergraduate research participation.