Laser trap measurements of flagellar membrane motility

Abstract

In addition to swimming motility, which is driven by propagation of bends along the flagellum, the unicellular green alga Chlamydomonas exhibits an unusual and alternative form of whole cell locomotion, called gliding motility. In gliding motility, a large flagellar membrane glycoprotein mediates flagellar membrane adhesion to solid substrates. This in turn activates a transmembrane signaling system that initiates the movement of a cross-linked cluster of glycoproteins within the plane of the flagellar membrane by activating and/or recruiting isoforms of the motor proteins kinesin and dynein. Flagellar membrane motility can be visualized through the bidirectional movement of microspheres adherent to the flagellar surface. This microsphere motility offers a unique, noninvasive experimental system for measuring the in vivo dynamics and regulation of microtubule-dependent molecular motors by using a laser trap transducer to capture and manipulate microspheres as they move along the flagellar surface. Detailed procedures for conducting such analyses are provided.