Biomedical subjects

Megan T Valentine

Publications and source records attributed to Megan T Valentine.

4 recordsLinked to original sources

PubMed2006

To step or not to step? How biochemistry and mechanics influence processivity in Kinesin and Eg5.

Conventional kinesin and Eg5 are essential nanoscale motor proteins. Single-molecule and presteady-state kinetic experiments indicate that both motors use similar strategies to generate movement along microtubules, despite having distinctly different in vivo functions. Single molecules of kinesin, a long-distance cargo transporter, are highly processive, binding the microtubule and taking 100 or more sequential steps at velocities of up to 700 nm/s before dissociating, whereas Eg5, a motor active in mitotic spindle assembly, is also processive, but takes fewer steps at a slower rate. By dissecting the structural, biochemical and mechanical features of these proteins, we hope to learn how kinesin and Eg5 are optimized for their specific biological tasks, while gaining insight into how biochemical energy is converted into mechanical work.

Animals↗

PubMed2006

Eg5 steps it up!

Understanding how molecular motors generate force and move microtubules in mitosis is essential to understanding the physical mechanism of cell division. Recent measurements have shown that one mitotic kinesin superfamily member, Eg5, is mechanically processive and capable of crosslinking and sliding microtubules in vitro. In this review, we highlight recent work that explores how Eg5 functions under load, with an emphasis on the nanomechanical properties of single enzymes.

Journal Article↗

PubMed2006

Individual dimers of the mitotic kinesin motor Eg5 step processively and support substantial loads in vitro.

Eg5, a member of the kinesin superfamily of microtubule-based motors, is essential for bipolar spindle assembly and maintenance during mitosis, yet little is known about the mechanisms by which it accomplishes these tasks. Here, we used an automated optical trapping apparatus in conjunction with a novel motility assay that employed chemically modified surfaces to probe the mechanochemistry of Eg5. Individual dimers, formed by a recombinant human construct Eg5-513-5His, stepped processively along microtubules in 8-nm increments, with short run lengths averaging approximately eight steps. By varying the applied load (with a force clamp) and the ATP concentration, we found that the velocity of Eg5 was slower and less sensitive to external load than that of conventional kinesin, possibly reflecting the distinct demands of spindle assembly as compared with vesicle transport. The Eg5-513-5His velocity data were described by a minimal, three-state model where a force-dependent transition follows nucleotide binding.

Adenosine Triphosphate↗

PubMed2003

Microscopic origin of light scattering in tissue.

A newly designed instrument, the static light-scattering (SLS) microscope, which combines light microscopy with SLS, enables us to characterize local light-scattering patterns of thin tissue sections. Each measurement is performed with an illumination beam of 70-microm diameter. On these length scales, tissue is not homogeneous. Both structural ordering and small heterogeneities contribute to the scattering signal. Raw SLS data consist of a two-dimensional intensity distribution map I(theta, phi), showing the dependence of the scattered intensity I on the scattering angle theta and the azimuthal angle phi. In contrast to the majority of experiments and to simulations that consider only the scattering angle, we additionally perform an analysis of the azimuthal dependence I(phi). We estimate different contributions to the azimuthal scattering variation and show that a significant fraction of the azimuthal amplitude is the result of tissue structure. As a demonstration of the importance of the structure-dependent part of the azimuthal signal, we show that this function of the scattered light alone can be used to classify tissue types with surprisingly high specificity and sensitivity.

Animals↗