6FFU

Solution NMR structure of CBM64 from S.thermophila using 20% 13C, 100% 15N

Summary for 6FFU

• Entry DOI: 10.2210/pdb6ffu/pdb
• NMR Information: BMRB: 34229
• Descriptor: Glycosyl hydrolase family 5 cellulase CBM64 (1 entity in total)
• Functional Keywords: cbm64, sugar binding protein
• Biological source: Spirochaeta thermophila
• Total number of polymer chains: 1
• Total formula weight: 10025.76

Authors

Heikkinen, H.A.,Iwai, H. (deposition date: 2018-01-09, release date: 2019-01-30, Last modification date: 2024-06-19)

Primary citation

Heikkinen, H.A.,Backlund, S.M.,Iwai, H.
NMR Structure Determinations of Small Proteins Using only One Fractionally 20% 13 C- and Uniformly 100% 15 N-Labeled Sample.
Molecules, 26:-, 2021

PubMed Abstract: Uniformly C- and N-labeled samples ensure fast and reliable nuclear magnetic resonance (NMR) assignments of proteins and are commonly used for structure elucidation by NMR. However, the preparation of uniformly labeled samples is a labor-intensive and expensive step. Reducing the portion of C-labeled glucose by a factor of five using a fractional 20% C- and 100% N-labeling scheme could lower the total chemical costs, yet retaining sufficient structural information of uniformly [C, N]-labeled sample as a result of the improved sensitivity of NMR instruments. Moreover, fractional C-labeling can facilitate reliable resonance assignments of sidechains because of the biosynthetic pathways of each amino-acid. Preparation of only one [20% C, 100% N]-labeled sample for small proteins (<15 kDa) could also eliminate redundant sample preparations of 100% N-labeled and uniformly 100% [C, N]-labeled samples of proteins. We determined the NMR structures of a small alpha-helical protein, the C domain of IgG-binding protein A from (SpaC), and a small beta-sheet protein, CBM64 module using [20% C, 100% N]-labeled sample and compared with the crystal structures and the NMR structures derived from the 100% [C, N]-labeled sample. Our results suggest that one [20% C, 100% N]-labeled sample of small proteins could be routinely used as an alternative to conventional 100% [C, N]-labeling for backbone resonance assignments, NMR structure determination, N-relaxation analysis, and ligand-protein interaction.

PubMed: 33535444
DOI: 10.3390/molecules26030747

Experimental method

SOLUTION NMR