Protein structures at fast MAS

Rapid data acquisition for de novo structure determination of proteins

We combine well-known techniques such as SIM-CP, co-evolution, bidirectional cross-polarization, orphan pathways and multiplexing, pioneered by several other groups in the field, to design proton-detected residue linking experiment. This sequence allows simultaneous acquisition of one new, CA(CON)CAH^Aand five previously known experiments NCAH^A, N(CO)CAH^A, CANH, CA(CO)NH and N(CACO)NH.⁴⁸For the microcrystalline ubiquitin sample, all these spectra could be recorded in about 1.7 days compared to about 5-7 days of measurement time required to independently acquire all the spectra.  These six spectra allow independent forward and backward backbone walks using the resolved 2D NH^Nand CAH^Aplanes. Rapid data acquisition combined with automated assignment makes this a compelling approach for de novo structural characterization of protein using nanomolar sample amounts.