Magic Angle Spinning Solid-State NMR Spectroscopy for Structural Studies of Protein Interfaces. Resonance Assignments of Differentially Enriched Escherichia coli Thioredoxin Reassembled by Fragment Complementation

• Published in: Journal of the American Chemical Society 2004-12, Vol.126 (50), p.16608-16620
• Main Authors: MARULANDA, Dabeiba, TASAYCO, Maria Luisa, MCDERMOTT, Ann, CATALDI, Marcela, ARRIARAN, Vilma, POLENOVA, Tatyana
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Carbon Isotopes; Escherichia coli Proteins - chemistry; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Models, Molecular; Molecular Sequence Data; Nitrogen Isotopes; Nuclear Magnetic Resonance, Biomolecular - methods; Peptide Fragments - chemistry; Polyethylene Glycols - chemistry; Proteins; Thioredoxins - chemistry
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja0464589

De novo site-specific backbone and side-chain resonance assignments are presented for U-15N(1-73)/U-13C,15N(74-108) reassembly of Escherichia coli thioredoxin by fragment complementation, determined using solid-state magic angle spinning NMR spectroscopy at 17.6 T. Backbone dihedral angles and secondary structure predicted from the statistical analysis of 13C and 15N chemical shifts are in general agreement with solution values for the intact full-length thioredoxin, confirming that the secondary structure is retained in the reassembled complex prepared as a poly(ethylene glycol) precipitate. The differential labeling of complementary thioredoxin fragments introduced in this work is expected to be beneficial for high-resolution structural studies of protein interfaces formed by protein assemblies by solid-state NMR spectroscopy.