RMPSHSSI · REVEALING MYOSIN'S POWER STROKE WITH HIGH-SPEED SCATTERING INTERFEROMETRY
FP7 — People (Marie Curie Actions)
- Duration
- 2013-10-01 → 2015-09-30
- EU contribution
- €221,606
- Participants
- 1
- Scheme
- MC-IEF
Lines connect the coordinator with its partners.
Results in brief
REVEALING MYOSIN'S POWER STROKE WITH HIGH-SPEED SCATTERING INTERFEROMETRY
Molecular motors are remarkable nanoscopic machines that are responsible for most forms of movement we encounter in the cellular world. One of their major roles relates to transport inside cells, which are highly organized and divided into many parts of different functions. This implies that cellular transport is required for efficient cellular function. In this work we visualized the movement of one of such proteins – myosin 5. One could think of myosin 5 as a small lorry, which can move remarkably long distances while caring a cargo. Remarkably, this protein resembles like a two legged creature that makes nanoscale steps. We used very small gold particles, smaller than myosin itself to label this protein. Myosin together with such a“gold tag” could be detected and its motion was recorded using a new microscopy technique called iSCAT (Interferometric Scattering Microscopy). iSCAT allowed us to ‘see’ the tiny steps of this motor; tens of nanometres small at up to 1000 frames per second. The results provided the first experimental evidence that this family of motor proteins take regular ‘stiff-legged’ steps. The findings are interesting to anyone trying to understand how the cellular machinery works, but it is also likely to be very important for those looking to build nanomachines. The capabilities of iSCAT for fast and very sensitive imaging have already attracted attention of several scientific groups working on different motors (kinesins and dyneins). With this new tool we can hope to understand how motor proteins work facilitating not only cellular transport but also cell division, cell replication or cell communication.
Data: CORDIS, © European Union
Project objective
The fundamental goal of microscopic imaging is to visualize and identify small objects and observe their motion. Of the many available techniques, single particle tracking has been an immensely powerful tool in the life sciences for studying the motion of individual objects and thereby the function of a large variety biological processes. Despite considerable advances, following the motion of individual molecules on the relevant time and size scale has remained an insurmountable challenge. Here, we propose to construct a novel optical microscope based on interferometric scattering (iSCAT) detection that will bridge the considerable gap between the temporal and spatial accuracy provided by current single molecule methods.The primary objective of this work is to utilize the unique imaging properties of interferometric scattering detection to directly visualize and thereby understand the power stroke of Myosin V. Myosin V is a molecular motor, a cargo transporter that moves along actin filaments while hydrolyzing ATP. A wealth of knowledge concerning myosin’s function has been acquired over the past decades. Nevertheless, many of the fundamental questions concerning the mechanism of the power stroke for these motors remain, largely due to fundamental limitations associated with the employed techniques. In this respect, the application of high-speed iSCAT to study the motion of Myosin V will represent a significant step forward in our understanding of the functionality of this important motor.We aim that this work will establish iSCAT as a viable and powerful in vitro imaging platform with capabilities that are several orders of magnitude beyond what is currently possible with state of-the-art single particle tracking approaches. As a consequence, this proposal will include specifics with respect both the biological and the technical challenges involved with the proposed studies.
Original text from CORDIS.
Participants
- THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD · OxfordCoordinatorUnited Kingdom
Links
Data: CORDIS, © European Union