NMR assignment of reduced form of copper, zinc superoxide dismutase from Salmonella enterica

Abstract

Almost complete assignment (97%) of NMR resonances was obtained for the reduced, Cu(I), form of prokaryotic CuZnSOD from Salmonella enterica. 13C direct detection was used to complement the standard bouquet of 1H detected triple resonance experiments and contributed to the identification of proline backbone resonances and to side chains assignments of Asx, Glx and aromatic rings. This is the only complete assignment available for monomer SOD from prokaryotic organisms.

Capturing a Reactive State of Amyloid Aggregates

“…The result shows an interesting contrast with 1 H solution NMR data for monomeric Cu ϩ -bound A␤(1-40), which display exchange broadening (Fig. 10) 15 N ␦ shift of 220 -227 ppm for His-82 and His-125 in ba3 oxidase subunit II, for which N ␦ sites of the histidine residues are coordinated to Cu ϩ (see Table 1, c) (85,86). Furthermore, the 13 C ␦ chemical shift is known to be very sensitive to the protonation or Cu ϩ -coordination state of the neighboring N ⑀ .…”

Toward Structural Dynamics: Protein Motions Viewed by Chemical Shift Modulations and Direct Detection of C′N Multiple-Quantum Relaxation

“…The enzymatic cavity is defined by an electrostatic loop, which contains a copper ion that can jump between oxidation states Cu 2+/+ and is responsible for the catalytic activity. The three-dimensional structure of this enzyme has been determined in solution, and the complete NMR assignment is available . Crystal structures of many homologous enzymes are also available …”

The Solution Structure of the Monomeric Copper, Zinc Superoxide Dismutase from Salmonella enterica: Structural Insights To Understand the Evolution toward the Dimeric Structure

“…Throughout this paper, we have used the amino acid numbering of the mature protein in which the first 19 residues have been deleted. Sequencespecific assignment of backbone and side chain resonances (22) was obtained using HSQC, HNCA, HNCO, HNCACB, CBCA(CO)NH,HBHA(CO)NH,(H)CCH-TOCSY,and 1 H, 15 N-NOESY-HSQC. In addition to the set of 2D and 3D 1 Hdetected experiments, we acquired 13 C direct detected CACO, CBCACO, CON and CC-COSY experiments (20,(29)(30)(31).…”

“…They confirmed proline sequential assignments and provided complete resonance assignment of aspartic acid/asparagine and glutamic acid/glutamine side chains and aromatic rings. Even if 13 C direct detection is especially useful for partially or completely unfolded proteins (32) or for systems affected by exchange broadening effects, CON experiments provide an alternative fingerprint also for a well-folded protein (22).…”

“…All 2D and 3D spectra were collected at 298 K on Bruker Avance 500, 600, 700, 800, and 900 MHz spectrometers, processed using the TOPSPIN software package (Bruker, Switzerland), and analyzed using CARA software (ETH, Zurich, Switzerland). NMR experiments required to obtain the complete assignment of protein resonances have been previously published (22) and are summarized in Supporting Information, Table S2, together with the NMR experiments used to obtain NOE intensities and 3 J coupling constants: 2D NOESY, 15 N-and 13 C-HSQC-NOESY, HNHA, and HNHB.…”