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. Author manuscript; available in PMC: 2021 Jun 14.
Published in final edited form as: Nat Struct Mol Biol. 2020 Dec 14;28(1):20–28. doi: 10.1038/s41594-020-00519-9

Fig. 3. Characterization of ciliopathy-based radial spoke head mutants.

Fig. 3.

a, Positions of mutations made in the recombinant Chlamydomonas head-neck complex are shown in spheres. The RS head surface is shown from the CP view, as indicated at the top right, and is transparent (not all mutations are on the head surface). RSP9 appears in two copies and thus, positions mutated in RSP9 are indicated twice. See Extended Data Fig. 6b–f for more details. b, SEC profiles of purified recombinant RS head variants, showing all mutants elute later than wild-type, except for Y244R in RSP9. c, Coomassie-stained SDS-PAGE of purified recombinant head variants. Black boxes indicate reduced amounts of protein compared to wild-type. Red box indicates elevated amounts of protein in RSP4 mutants, which is likely a chaperone. Uncropped gel image is shown in Supplementary Figure 1. d, Representative negative-stain EM class averages for recombinant RS head variants. Box size is 4.4 nm. e, Representative melting curves for recombinant RS head variants, showing all mutants denature at a lower temperature than WT. Apparent denaturation temperatures are reported in Table 2.