KIF18A sNL variant is less processive but faster than WT KIF18A and more hindered by obstacles. KIF18A sNL variant is less processive but faster than WT.

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KIF18A sNL variant is less processive but faster than WT KIF18A and more hindered by obstacles. KIF18A sNL variant is less processive but faster than WT KIF18A and more hindered by obstacles. (A) Cartoon depiction of single-molecule TIRF assay setup without obstacles on microtubules. (B) Picture of single rhodamine (Rh)-labeled MT in single-molecule TIRF (top); representative kymographs of GFP-KIF18A1–480 WT (WT) and GFP-KIF18A1–480 sNL1 (sNL1) processive events. (C) Histogram distributions of run lengths (top) and velocities (bottom) of processive events observed. P < 0.0001 for both run length and velocity comparing WT and sNL1 by unpaired t test. (D) Cartoon depiction of single-molecule TIRF assay set up with Alexa 647 3RS tau as an obstacle on MT. (E) Picture of single Alexa 647 3RS tau–coated MT (top); representative kymographs of GFP-KIF18A1–480 WT (WT) and GFP-KIF18A1–480 sNL1 (sNL1) processive events on tau-coated MT (bottom). (F) Histogram distributions of run lengths (top) and velocities (bottom) of processive events observed on tau-coated MT. Statistical significance comparing no tau versus tau by unpaired t test: run length, WT P = 0.0006; sNL1 P < 0.0001; velocity, WT P = 0.0391; and sNL1 P = 0.6170. Data obtained from two independent protein preps and 16 individual TIRF assays with the following total processive events: no tau: WT (190), sNL1 (214); with tau: WT (194), sNL1 (207). MT, microtubules. Source data are available for this figure. Heidi LH Malaby et al. LSA 2019;2:e201800169 © 2019 Malaby et al.