Electron Spin Density on the Axial His Ligand of High-spin and Low-spin Nitrophorin 2 Probed by Heteronuclear NMR Spectroscopy

Overview

Journal Inorg Chem

Specialty Chemistry

Date 2013 Jan 19

PMID 23327568

Citations 4

Authors

Luciano A Abriata

Maria-Eugenia Zaballa

Robert E Berry

Fei Yang

Hongjun Zhang

F Ann Walker

Alejandro J Vila

Affiliations

Soon will be listed here.

Abstract

The electronic structure of heme proteins is exquisitely tuned by the interaction of the iron center with the axial ligands. NMR studies of paramagnetic heme systems have been focused on the heme signals, but signals from the axial ligands have been rather difficult to detect and assign. We report an extensive assignment of the (1)H, (13)C and (15)N resonances of the axial His ligand in the NO-carrying protein nitrophorin 2 (NP2) in the paramagnetic high-spin and low-spin forms, as well as in the diamagnetic NO complex. We find that the high-spin protein has σ spin delocalization to all atoms in the axial His57, which decreases in size as the number of bonds between Fe(III) and the atom in question increases, except that within the His57 imidazole ring the contact shifts are a balance between positive σ and negative π contributions. In contrast, the low-spin protein has π spin delocalization to all atoms of the imidazole ring. Our strategy, adequately combined with a selective residue labeling scheme, represents a straightforward characterization of the electron spin density in heme axial ligands.

Citing Articles

Dimerization of nitrophorin 4 at low pH and comparison to the K1A mutant of nitrophorin 1.

Berry R, Yang F, Shokhireva T, Amoia A, Garrett S, Goren A Biochemistry. 2014; 54(2):208-20.

PMID: 25489673 PMC: 4303305. DOI: 10.1021/bi5013047.

NMR studies of the dynamics of high-spin nitrophorins: comparative studies of NP4 and NP2 at close to physiological pH.

Berry R, Muthu D, Yang F, Walker F Biochemistry. 2014; 54(2):221-39.

PMID: 25486224 PMC: 4303294. DOI: 10.1021/bi501305a.

NMR investigations of nitrophorin 2 belt side chain effects on heme orientation and seating of native N-terminus NP2 and NP2(D1A).

Berry R, Muthu D, Shokhireva T, Garrett S, Goren A, Zhang H J Biol Inorg Chem. 2013; 19(4-5):577-93.

PMID: 24292244 PMC: 4031258. DOI: 10.1007/s00775-013-1063-8.

NMR studies of the dynamics of nitrophorin 2 bound to nitric oxide.

Muthu D, Berry R, Zhang H, Walker F Biochemistry. 2013; 52(45):7910-25.

PMID: 24116947 PMC: 3947638. DOI: 10.1021/bi4010396.

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