Our pipeline consists of generating different constructs allowing the expression of a protein of interest. We then use NMR spectroscopy to select constructs that produce properly folded protein before setting crystal trays.
2D [1H, 15N] TROSY-HSQC spectrum of eIF4E (A) and 4E-BP144-87 (B) in the bound form acquired on a 900 MHz spectrometer.
Adapted from Sekiyama, N., et al. (2015). Proc Natl Acad Sci USA. 112: E4036-E4045.
Cartoon representation of the Leishmania major IF4E-1/4E-IP11-52 complex (side view and dorsal view, PDB ID: 5WB5), where residues 13 to 210 of LmIF4E-1 and residues 4 to 43 of Lm4E-IP11-52 are shown. LmIF4E-1 and Lm4E-IP1 are shown in cyan and purple, respectively.
Adapted from Meleppattu, S., et al. (2018). Nucleic Acids Research, 46: 3791-3801.