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. Author manuscript; available in PMC: 2021 Apr 21.
Published in final edited form as: Biochemistry. 2020 Apr 13;59(15):1527–1536. doi: 10.1021/acs.biochem.0c00094

Figure 4.

Figure 4.

Skp1ΔF97E is a stable and functional monomer in solution. (A) c(s) distributions of 100 µM Skp1Δ or Skp1ΔF97E are shown in cyan and black, respectively. (B) 1H/15N-HSQC of 100 µM Skp1ΔF97E at 900 MHz and 35°C, with a 4 h collection time. The 500 μM spectrum (not shown) was indistinguishable. (C, D) Skp1ΔF97E binds the model F-box protein Fbs1. His6Fbs1 (1.5 μM) and an estimated 2.25 µM Skp1Δ (C) or Skp1Δ(F97E) (D) were analyzed on a Superdex 200 gel filtration column. Elution was monitored by A280, which favors detection of Fbs1 relative to Skp1 because of its higher extinction coefficient.