DOI: 10.5061/dryad.1rn8pk12h
title: Data from: Heterogeneous distribution of kinesin-streptavidin complexes revealed by mass photometry

description: Kinesin-streptavidin complexes are widely used in microtubule-based
 active-matter studies. The stoichiometry of the complexes is
 empirically tuned but experimentally challenging to determine.
 Here, mass photometry measurements reveal heterogenous
 distributions of kinesin-streptavidin complexes. Our binding
 model indicates that heterogeneity arises from both the
 kinesin-streptavidin mixing ratio and the kinesin-biotinylation
 efficiency.
description: # Data from: Heterogeneous distribution of kinesin-streptavidin complexes
 revealed by Mass Photometry
 [https://doi.org/10.5061/dryad.1rn8pk12h](https://doi.org/10.5061/dryad.1rn8pk12h) Data underlying Figs. 2-4 and Figs. S1-S10. ## Description of the data and file structure Data underlying Figs. 2-4, Fig. S1, Figs. S2b-S6b, and Figs. S7b-S10 are provided in .xlsx file format. Data underlying Figs. S2a and S7a  are provided in .tif file format. Details below: * **Figure 2.xls:**  data underlying Fig. 2 in the main text. This file contains three tabs. * MassPhotometry Counts Tab: * Column A: Mass (kDa) * Column B: Counts of Kinesin (K) * Column C: Counts of Streptavidin (S) * Column D: Counts of K:S 0.4:1 * Column E: Counts of K:S 0.8:1 * Column F: Counts of K:S 1.6:1 * Column G: Counts of K:S 2.2:1 * Column H: Counts of K:S 3.0:1 * Column I: Counts of K:S 3.6:1 * Global Fit (red lines) Tab:  red lines in Fig. 2 * Columns A and B: Mass (kDa) and Best-fit to Counts of Kinesin (K) * Column D and E: Mass (kDa) and Best-fit to  Counts of Streptavidin (S) * Column G and H: Mass (kDa) and Best-fit to  Counts of K:S 0.4:1 * Column J and K: Mass (kDa) and Best-fit to  Counts of K:S 0.8:1 * Column M and N: Mass (kDa) and Best-fit to  Counts of K:S 1.6:1 * Column P and Q: Mass (kDa) and Best-fit to  Counts of K:S 2.2:1 * Column S and T: Mass (kDa) and Best-fit to  Counts of K:S 3.0:1 * Column V and W: Mass (kDa) and Best-fit to  Counts of K:S 3.6:1 * Individual Fit (black lines) Tab: black lines in Figure 2, right panel. * Columns A and B: Mass (kDa) and Best-fit for peak iii in K:S 0.4:1 * Columns D, E, and F: Mass (kDa) and Best-fits for peaks iii and iv in K:S 0.8:1 * Columns H, I, and J: Mass (kDa) and Best-fits for peaks iii and iv in K:S 1.6:1 * Columns L, M, and N: Mass (kDa) and Best-fits for peaks iv and v in K:S 2.2:1 * Columns P, Q, and R: Mass (kDa) and Best-fits for peaks iv and v in K:S 3.0:1 * Columns T, U, and V: Mass (kDa) and Best-fits for peaks iv and v in K:S 3.6:1 * **Figure 3.xlsx:** data underlying Fig. 3 in the main text. This file contains two tabs. * Scatter Data Tab: * Column A:  Expected mass (kDa) * Column B:  Measured mass, mean value (kDa) * Column C:  Measured mass, standard deviation (kDa) * Linear Fit Tab:  red line in Fig. 3 * Column A: Expected mass (kDa) * Column B: Best-fit to measured mass (kDa) * **Figure 4.xlsx:** data underlying Fig. 4 in the main text. This file contains three tabs. * Scatter Data Tab: * Column A: Kinesin-streptavidin mixing ratio * Column B: Relative abundance of 1-kinesin complex (%) * Column C:  Error bar in the relative abundance of 1-kinesin complex (%) * Column D: Relative abundance of 2-kinesin complex (%) * Column E:  Error bar in the relative abundance of 2-kinesin complex (%) * Column F: Relative abundance of 3-kinesin complex (%) * Column G:  Error bar in the relative abundance of 3-kinesin complex (%) * Column H: Relative abundance of higher-order complex (%) * Column I:  Error bar in the relative abundance of higher-order complex (%) * BindingModel_DashedLines Tab: dashed lines in Fig. 4 * Column A: Kinesin-streptavidin mixing ratio * Column B:  Relative abundance of 1-kinesin complex (%) * Column C:  Relative abundance of 2-kinesin complex (%) * Column D:  Relative abundance of 3-kinesin complex (%) * Column E:  Relative abundance of 4-kinesin complex (%) * BindingModel_BlueLines Tab: blue lines in Fig. 4 * Column A: Kinesin-streptavidin mixing ratio * Column B:  Relative abundance of 1-kinesin complex (%) * Column C:  Relative abundance of 2-kinesin complex (%) * Column D:  Relative abundance of 3-kinesin complex (%) * Column E:  Relative abundance of 4-kinesin complex (%) * **Figure S1.xlsx:** data underlying Fig. S1 in the supplement. This file contains two tabs. * ScatterData Tab: * Column A: Mass (kDa) * Column B: Interference intensity (a.u.) * LinearFit Tab: black line in Fig. S1 * Column A: Mass (kDa) * Column B: Best-fit to interference intensity (a.u.) * **Figure S2a.tif:**  gel file for Fig. S2a in the supplement. * **Figure S2b.xlsx:** data underlying Fig. S2 in the supplement. This file contains two tabs. * ScatterData Tab: * Column A: Kinesin dilution relative to Lane 2 * Column B: Kinesin-streptavidin mixing ratio * LinearFit Tab: black line in Fig. S2b * Column A: Kinesin dilution relative to Lane 2 * Column B: Best-fit to kinesin-streptavidin mixing ratio * **Figure S3a.xlsx:** data underlying Fig. S3a in the supplement. This file contains one tab. * Counts Tab: * Column A: Mass (kDa) * Column B: Counts for Measurements 1-3 * Column C: Counts for Measurements 4-6 * Column D: Counts for Measurements 7-9 * Column E: Counts for Measurements 10-12 * Column F: Mean counts for Measurements 1-12 * Column G: Standard deviation in the counts of Measurements 1-12 * **Figure S3b.xlsx:** data underlying Fig. S3b in the supplement. This file contains two tabs. * Left panel Tab: data for the left panel * Column A: Mass (kDa) * Column B: Counts for Sample 1 * Column C: Counts for Sample 2 * Column D: Mean counts of Sample 1 and 2 * Column E: Standard deviation of Sample 1 and 2 * Right panel Tab: data for the right panel * Column A: Mass (kDa) * Column B: Normalized counts of Sample 1 (%) * Column C: Normalized counts of Sample 2 (%) * Column D: Mean of the normalized counts of Sample 1 and Sample 2 (%) * Column E: Standard deviation of the normalized counts of Sample 1 and Sample 2 (%) * **Figure S4.xlsx:** data underlying Fig. S4 in the supplement. This file contains one tab. * MassPhotometry Counts Tab: * Column A: Mass (kDa) * Column B: Counts of Kinesin (K) * Column C: Counts of Streptavidin (S) * Column D: Counts of K:S 0.4:1 * Column E: Counts of K:S 0.8:1 * Column F: Counts of K:S 1.6:1 * Column G: Counts of K:S 2.2:1 * Column H: Counts of K:S 3.0:1 * Column I: Counts of K:S 3.6:1 * **Figure S5.xlsx:** data underlying Fig. S5 in the supplement. This file contains one tab. * Counts Tab: * Column A: Mass (kDa) * Column B: Counts of K:S 0.8:1 * Column C: Counts of K:S 3.0:1 * **Figure S6a.xlsx:** data underlying Fig. S6a in the supplement. This file contains three tabs. * Counts Tab: * Column A: Mass (kDa) * Column B: Counts of Sample 1 * Column C: Counts of Sample 2 * Sample 1 Fits Tab: best-fits to Sample 1 data * Column A and B: Mass (kDa) and Best-fit for Peak ii * Column D and E: Mass (kDa) and Best-fit for Peak iii * Column G and H: Mass (kDa) and Best-fit for Peak iv * Column J and K: Mass (kDa) and Best-fit for Peak v * Sample 2 Fits Tab: best-fits to Sample 2 data * Column A and B: Mass (kDa) and Best-fit for Peak ii * Column D and E: Mass (kDa) and Best-fit for Peak iii * Column G and H: Mass (kDa) and Best-fit for Peak iv * Column J and K: Mass (kDa) and Best-fit for Peak v * **Figure S6b.xlsx:** data underlying Fig. S6b in the supplement. This file contains one tab. * Column A: Kinesin-streptavidin complex species * Column B: Relative abundance of complex species for Sample 1 (%) * Column C: Relative abundance of complex species for Sample 2 (%) * **Figure S7a_kinesin.tif:** gel file for Fig. S7a (kinesin) in the supplement. * **Figure S7a_streptavidin.tif:** gel file for Fig. S7a (streptavidin) in the supplement. * **Figure S7b.xlsx:** data underlying Fig. S7b in the supplement. This file contains one tab. * Counts Tab: * Column A: Deconvoluted mass (Da) * Column B: Counts * **Figure S8.xlsx:** data underlying Fig. S8 in the supplement. This file contains one tab. * Sheet1 Tab: * Column A & Rows 2-102: Overall scaling for mixing ratio * Column B-AF & Row 1: % kinesin biotinylation * Column B-AF & Rows 2-102: Reduced 𝜒2 for each combination of the % kinesin-biotinylation efficiency and the overall scaling for the kinesin-streptavidin mixing ratio * **Figure S9.xlsx:** data underlying Fig. S9 in the supplement. This file contains one tab. * Sheet1 Tab: * Column A: Streptavidin concentration (uM) * Column B: Relative abundance of 1-kinesin complex (%) * Column C: Error bar in the relative abundance of 1-kinesin complex (%) * Column D: Relative abundance of 2-kinesin complex (%) * Column E: Error bar in the relative abundance of 2-kinesin complex (%) * Column F: Relative abundance of 3-kinesin complex (%) * Column G: Error bar in the relative abundance of 3-kinesin complex (%) * Column H: Relative abundance of higher-order complex (%) * Column I: Error bar in the relative abundance of higher-order complex (%) * **Figure S10.xlsx:** data underlying Fig. S10 in the supplement. This file contains five tabs. * 100% biotinylation Tab: * Column A: kinein-streptavidin mixing ratio * Column B: Relative abundance of 1-kinesin complex (%) * Column C: Relative abundance of 2-kinesin complex (%) * Column D: Relative abundance of 3-kinesin complex (%) * Column E: Relative abundance of 4-kinesin complex (%) * 80% biotinylation Tab: * Column A: kinein-streptavidin mixing ratio * Column B: Relative abundance of 1-kinesin complex (%) * Column C: Relative abundance of 2-kinesin complex (%) * Column D: Relative abundance of 3-kinesin complex (%) * Column E: Relative abundance of 4-kinesin complex (%) * 70% biotinylation Tab: * Column A: kinein-streptavidin mixing ratio * Column B: Relative abundance of 1-kinesin complex (%) * Column C: Relative abundance of 2-kinesin complex (%) * Column D: Relative abundance of 3-kinesin complex (%) * Column E: Relative abundance of 4-kinesin complex (%) * 60% biotinylation Tab: * Column A: kinein-streptavidin mixing ratio * Column B: Relative abundance of 1-kinesin complex (%) * Column C: Relative abundance of 2-kinesin complex (%) * Column D: Relative abundance of 3-kinesin complex (%) * Column E: Relative abundance of 4-kinesin complex (%) * 50% biotinylation Tab: * Column A: kinein-streptavidin mixing ratio * Column B: Relative abundance of 1-kinesin complex (%) * Column C: Relative abundance of 2-kinesin complex (%) * Column D: Relative abundance of 3-kinesin complex (%) * Column E: Relative abundance of 4-kinesin complex (%)