Characterization of AlkL Protein via NMR

Solution NMR spectra were recorded at ω_0H/2π = 1 GHz and 600 MHz on Bruker Avance III instruments equipped with cryogenic probes using ²H,¹³C,¹⁵N-labeled AlkL. Solid-state NMR spectra were recorded at ω_0H/2π = 1 GHz and 800 MHz, on Bruker Avance III instruments equipped with 0.7- and 1.3-mm HCN probes spinning at MAS rates of 111.111 kHz (¹³C,¹⁵N-labeled AlkL) or 60 kHz (²H,¹³C,¹⁵N-labeled AlkL). Spectrometer settings, as well as acquisition parameters specific for each 3D and 4D spectrum, and for site-specific ¹⁵N R₁ (26 (link)) and (R_1ρ) (36 (link)) rates, are discussed in SI Appendix, Supplementary Text and summarized in SI Appendix, Table S1. The spectral quality underlying resonance assignment datasets (37 (link), 38 (link)) is demonstrated in the pair of CA-N-H correlation spectra shown in SI Appendix, Fig. S13 and the assignments are discussed in detail in a separate manuscript (39 (link)). CSPs induced by the presence of carvone or octane were obtained from (H)CANH and (H)CONH spectra, and were calculated as the RMSD of ¹H, ¹³CA, and ¹⁵N shifts, with relative scaling of 1, 0.3, and 0.15, respectively (40 ).