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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.

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Speed versus assay time for four types of casein passivation.Black circles are alpha casein passivation, red squares are beta casein                            passivation, green up pointing triangles are whole casein passivation,                            and blue down pointing triangles are mixed casein passivation. Each data                            point is the mean from three different samples, taken at approximately                            the same assay time. Error bars represent the standard error of the                            mean. Alpha casein had the most consistent average speed measurements at                            949±4 nm/s. Whole casein and mixed casein averaged to                            966±7 nm/s and 966±6 nm/s respectively. Beta casein                            averaged to 870±30 nm/s.
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pone-0019522-g003: Speed versus assay time for four types of casein passivation.Black circles are alpha casein passivation, red squares are beta casein passivation, green up pointing triangles are whole casein passivation, and blue down pointing triangles are mixed casein passivation. Each data point is the mean from three different samples, taken at approximately the same assay time. Error bars represent the standard error of the mean. Alpha casein had the most consistent average speed measurements at 949±4 nm/s. Whole casein and mixed casein averaged to 966±7 nm/s and 966±6 nm/s respectively. Beta casein averaged to 870±30 nm/s.

Mentions: Figure 3 shows the mean speed measurements for 15 different regions of interest for the alpha, beta, whole and mixed casein assays. Each data point is the mean of a region of interest with SEM from three separate samples. The passivator that gave the most consistent speed was alpha casein. The mean speed and SEM from our alpha casein measurement was 949±4 nm/s. Purchased whole casein and mixed casein performed remarkably similarly and displayed average speed values of 966±7 nm/s and 966±7 nm/s respectively. Bovine beta casein performed poorly in comparison to alpha, whole, or mixed caseins and we measured the mean speed to be 870±30 nm/s. Figure 4 shows the observed speeds for kappa casein passivation. Since there were so many areas where no motility was observed in this assay, it was difficult to determine a mean speed measurement for each assay as was done in Figure 3. However, it does appear that when motile, the speeds were around 870–880 nm/s with kappa casein as the surface passivator. This was similar to how beta casein performed.


Effects of surface passivation on gliding motility assays.

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

Speed versus assay time for four types of casein passivation.Black circles are alpha casein passivation, red squares are beta casein                            passivation, green up pointing triangles are whole casein passivation,                            and blue down pointing triangles are mixed casein passivation. Each data                            point is the mean from three different samples, taken at approximately                            the same assay time. Error bars represent the standard error of the                            mean. Alpha casein had the most consistent average speed measurements at                            949±4 nm/s. Whole casein and mixed casein averaged to                            966±7 nm/s and 966±6 nm/s respectively. Beta casein                            averaged to 870±30 nm/s.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0019522-g003: Speed versus assay time for four types of casein passivation.Black circles are alpha casein passivation, red squares are beta casein passivation, green up pointing triangles are whole casein passivation, and blue down pointing triangles are mixed casein passivation. Each data point is the mean from three different samples, taken at approximately the same assay time. Error bars represent the standard error of the mean. Alpha casein had the most consistent average speed measurements at 949±4 nm/s. Whole casein and mixed casein averaged to 966±7 nm/s and 966±6 nm/s respectively. Beta casein averaged to 870±30 nm/s.
Mentions: Figure 3 shows the mean speed measurements for 15 different regions of interest for the alpha, beta, whole and mixed casein assays. Each data point is the mean of a region of interest with SEM from three separate samples. The passivator that gave the most consistent speed was alpha casein. The mean speed and SEM from our alpha casein measurement was 949±4 nm/s. Purchased whole casein and mixed casein performed remarkably similarly and displayed average speed values of 966±7 nm/s and 966±7 nm/s respectively. Bovine beta casein performed poorly in comparison to alpha, whole, or mixed caseins and we measured the mean speed to be 870±30 nm/s. Figure 4 shows the observed speeds for kappa casein passivation. Since there were so many areas where no motility was observed in this assay, it was difficult to determine a mean speed measurement for each assay as was done in Figure 3. However, it does appear that when motile, the speeds were around 870–880 nm/s with kappa casein as the surface passivator. This was similar to how beta casein performed.

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