Table 1.
MS/MS analysis results for in-solution digestion of RBD obtained from HEK-293T and from P. pastoris.
| Sample | MW(expt) (Da) | MW(calc) (Da) | ppm | Peptide | Protein sequence |
|---|---|---|---|---|---|
| HEK-293T | 989.48 | 989.52 | − 47.17 | K.SNLKPFER.D | SRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKLPETGHHHHHH |
| 1112.49 | 1112.54 | − 42.78 | R.FASVYAWNR.K | ||
| 1217.53 | 1217.58 | − 41.21 | K.VGGNYNYLYR.L | ||
| 2045.92 | 2045.99 | − 31.94 | K.LNDLCFTNVYADSFVIR.G + Carbamidomethyl (C) | ||
| 2264.98 | 2265.04 | − 24.05 | K.LPDDFTGCVIAWNSNNLDSK.V + Carbamidomethyl (C) | ||
| 2372.04 | 2372.10 | − 23.92 | R.ISNCVADYSVLYNSASFSTFK.C + Carbamidomethyl (C) | ||
| P. pastoris | 1027.51 | 1027.57 | − 52.05 | R.VQPTESIVR.F | RVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKLPETGHHHHHH |
| 1217.52 | 1217.58 | − 49.42 | K.VGGNYNYLYR.L | ||
| 2045.89 | 2045.99 | − 46.60 | K.LNDLCFTNVYADSFVIR.G + Carbamidomethyl (C) |
Protein coverage was of ∼ 40 and ∼ 60%, for RBD produced in P. pastoris and HEK-293T cells, respectively. The experimentally obtained (MW (expt)) and the calculated (MW (calc)) molecular weights of the peptides, and their difference (ppM) are shown. The protein sequence of each construct is shown, bold indicate the peptides identified either by MS/MS analysis or peptide mass fingerprint (less than 60 ppm error).