13C- 13C NOESY spectra of a 480 kDa protein: solution NMR of ferritin.
Molecular size has limited solution NMR analyses of proteins. We report (13)C-(13)C NOESY experiments on a 480 kDa protein, the multi-subunit ferritin nanocage with gated pores. By exploiting (13)C-resonance-specific chemical shifts and spin diffusion effects, we identified 75% of the amino acids, with intraresidue C-C connectivities between nuclei separated by 1-4 bonds. These results show the potential of (13)C-(13)C NOESY for solution studies of molecular assemblies >100 kDa.
Matzapetakis
Manolis
M
Department of Chemistry, CERM, University of Florence, Via Luigi Sacconi, 6, Sesto Fiorentino, FI, 50019, Italy.
Turano
Paola
P
Theil
Elizabeth C
EC
Bertini
Ivano
I
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002244:Carbon / Q000032:analysis* / Q000737:chemistry; D002247:Carbon Isotopes; D005293:Ferritins / Q000737:chemistry*; D008970:Molecular Weight; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017433:Protein Structure, Secondary; D011892:Rana catesbeiana; D012996:Solutions; D013997:Time Factors
D000596:Amino Acids; D002247:Carbon Isotopes; D012996:Solutions; D002244:Carbon; D005293:Ferritins