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Effects of surface passivation on gliding motility assays.

Maloney A, Herskowitz LJ, Koch SJ - PLoS ONE (2011)

Bottom Line: Beta casein did not support motility very well and averaged speeds of 870±30 nm/s.Kappa casein supported motility very poorly and we were unable to obtain an average speed.Finally, we observed that mixing alpha, beta, and kappa casein with the proportions found in bovine whole casein supported motility and averaged speeds of 966±6 nm/s.

View Article: PubMed Central - PubMed

Affiliation: Department of Physics and Astronomy and Center for High Technology Materials, University of New Mexico, Albuquerque, New Mexico, United States of America. amaloney@unm.edu

ABSTRACT
In this study, we report differences in the observed gliding speed of microtubules dependent on the choice of bovine casein used as a surface passivator. We observed differences in both speed and support of microtubules in each of the assays. Whole casein, comprised of α(s1), α(s2), β, and κ casein, supported motility and averaged speeds of 966±7 nm/s. Alpha casein can be purchased as a combination of α(s1) and α(s2) and supported gliding motility and average speeds of 949±4 nm/s. Beta casein did not support motility very well and averaged speeds of 870±30 nm/s. Kappa casein supported motility very poorly and we were unable to obtain an average speed. Finally, we observed that mixing alpha, beta, and kappa casein with the proportions found in bovine whole casein supported motility and averaged speeds of 966±6 nm/s.

Show MeSH
Observations of microtubules in flow cells passivated by different                            types of casein.Images have been false colored with ImageJ's Green Fire Blue LUT and                            have had dead pixels removed using an interpolation function.                                A. Alpha casein (Sigma C6780) passivation showed                            support for both long and short microtubules. B. Beta                            casein (Sigma C6095) passivation did not support motility very well and                            did not support shorter microtubules. C. Kappa casein                            (Sigma C0406) did support motility but only in limited regions of the                            flow cell. In other regions near the center of the channel                                (D) the microtubules were stuck to the slide and did                            not exhibit motility. E. Commercially available whole                            casein (Sigma C7078) is the standard for surface passivation for the                            gliding motility assay. It supported both long and very short                            microtubules. F. Mixed casein which was made from                            49% α, 37% β, and 14% κ casein worked                            just as well as alpha casein and whole casein passivation in terms of                            number and sizes of microtubules exhibiting motility.
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pone-0019522-g001: Observations of microtubules in flow cells passivated by different types of casein.Images have been false colored with ImageJ's Green Fire Blue LUT and have had dead pixels removed using an interpolation function. A. Alpha casein (Sigma C6780) passivation showed support for both long and short microtubules. B. Beta casein (Sigma C6095) passivation did not support motility very well and did not support shorter microtubules. C. Kappa casein (Sigma C0406) did support motility but only in limited regions of the flow cell. In other regions near the center of the channel (D) the microtubules were stuck to the slide and did not exhibit motility. E. Commercially available whole casein (Sigma C7078) is the standard for surface passivation for the gliding motility assay. It supported both long and very short microtubules. F. Mixed casein which was made from 49% α, 37% β, and 14% κ casein worked just as well as alpha casein and whole casein passivation in terms of number and sizes of microtubules exhibiting motility.

Mentions: Bovine alpha casein (a mixture of αs1 37% and αs2 10%) constitutes approximately 47% of whole bovine casein [20]. This passivation was capable of supporting small microtubules and longer ones as can be seen in Figure 1A. When using alpha casein, the gliding motility assay worked every time except when we deemed the kinesin or antifade system to have lost its effectiveness for maintaining a gliding assay.


Effects of surface passivation on gliding motility assays.

Maloney A, Herskowitz LJ, Koch SJ - PLoS ONE (2011)

Observations of microtubules in flow cells passivated by different                            types of casein.Images have been false colored with ImageJ's Green Fire Blue LUT and                            have had dead pixels removed using an interpolation function.                                A. Alpha casein (Sigma C6780) passivation showed                            support for both long and short microtubules. B. Beta                            casein (Sigma C6095) passivation did not support motility very well and                            did not support shorter microtubules. C. Kappa casein                            (Sigma C0406) did support motility but only in limited regions of the                            flow cell. In other regions near the center of the channel                                (D) the microtubules were stuck to the slide and did                            not exhibit motility. E. Commercially available whole                            casein (Sigma C7078) is the standard for surface passivation for the                            gliding motility assay. It supported both long and very short                            microtubules. F. Mixed casein which was made from                            49% α, 37% β, and 14% κ casein worked                            just as well as alpha casein and whole casein passivation in terms of                            number and sizes of microtubules exhibiting motility.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3108588&req=5

pone-0019522-g001: Observations of microtubules in flow cells passivated by different types of casein.Images have been false colored with ImageJ's Green Fire Blue LUT and have had dead pixels removed using an interpolation function. A. Alpha casein (Sigma C6780) passivation showed support for both long and short microtubules. B. Beta casein (Sigma C6095) passivation did not support motility very well and did not support shorter microtubules. C. Kappa casein (Sigma C0406) did support motility but only in limited regions of the flow cell. In other regions near the center of the channel (D) the microtubules were stuck to the slide and did not exhibit motility. E. Commercially available whole casein (Sigma C7078) is the standard for surface passivation for the gliding motility assay. It supported both long and very short microtubules. F. Mixed casein which was made from 49% α, 37% β, and 14% κ casein worked just as well as alpha casein and whole casein passivation in terms of number and sizes of microtubules exhibiting motility.
Mentions: Bovine alpha casein (a mixture of αs1 37% and αs2 10%) constitutes approximately 47% of whole bovine casein [20]. This passivation was capable of supporting small microtubules and longer ones as can be seen in Figure 1A. When using alpha casein, the gliding motility assay worked every time except when we deemed the kinesin or antifade system to have lost its effectiveness for maintaining a gliding assay.

Bottom Line: Beta casein did not support motility very well and averaged speeds of 870±30 nm/s.Kappa casein supported motility very poorly and we were unable to obtain an average speed.Finally, we observed that mixing alpha, beta, and kappa casein with the proportions found in bovine whole casein supported motility and averaged speeds of 966±6 nm/s.

View Article: PubMed Central - PubMed

Affiliation: Department of Physics and Astronomy and Center for High Technology Materials, University of New Mexico, Albuquerque, New Mexico, United States of America. amaloney@unm.edu

ABSTRACT
In this study, we report differences in the observed gliding speed of microtubules dependent on the choice of bovine casein used as a surface passivator. We observed differences in both speed and support of microtubules in each of the assays. Whole casein, comprised of α(s1), α(s2), β, and κ casein, supported motility and averaged speeds of 966±7 nm/s. Alpha casein can be purchased as a combination of α(s1) and α(s2) and supported gliding motility and average speeds of 949±4 nm/s. Beta casein did not support motility very well and averaged speeds of 870±30 nm/s. Kappa casein supported motility very poorly and we were unable to obtain an average speed. Finally, we observed that mixing alpha, beta, and kappa casein with the proportions found in bovine whole casein supported motility and averaged speeds of 966±6 nm/s.

Show MeSH