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. 2018 Mar 7;13(3):e0193947. doi: 10.1371/journal.pone.0193947

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© 2018 Lim et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 3 — Representative EPR-DEER data of GCAP1^CL(E57C) (A), GCAP1^CL(E133C) (C), and GCAP1^CL(E154C) (E), and corresponding distance distributions of GCAP1^CL(E57C) (B), GCAP1^CL(E133C) (D), and GCAP1^CL(E154C) (F). GCAP1 samples were in the Ca²⁺-free/Mg²⁺-bound state. Similar DEER data were observed for GCAP1 in the Ca²⁺-bound state. A nitroxide spin-label (MTSSL) was covalently attached to the sole Cys residue in each mutant. The distance distributions and average intermolecular distances were calculated on the basis of the DEER data as described in Methods. The bimodal distance distribution for GCAP1^CL(E57C) suggest there are two populations of GCAP1 dimers with distinct structure near E57C. The DEER intermolecular distances were measured to be 26±3/50±5 Å (GCAP1^CL(E57C) in panel B), 50 ±5 Å (GCAP1^CL(E133C) in panel D), and 28 ±3 Å (GCAP1^CL(E154C) in panel F).