NMR studies of lysozyme surface accessibility by using different paramagnetic relaxation probes.
Paramagnetic probes, whose approach to proteins can be monitored by nuclear magnetic resonance (NMR) studies, have been found to be of primary relevance for investigating protein surfaces' accessibility. Here, a Gd(III) neutral complex which contains two metal ions, [Gd2(L7)(H2O)2], is suggested as a paramagnetic probe particularly suited for systematic NMR investigation of protein surface accessibility, due to an expected high relaxivity and to the lack of electric charge which could favor specific interactions. Hen egg white lysozyme has been used as a model system to verify the absence of preferential approaches of this paramagnetic probe to specific protein moieties by comparing paramagnetic perturbation profiles of 1H-13C HSQC signals obtained in the presence of TEMPOL and [Gd2(L7)(H2O)2]. From the similarity of the measured paramagnetic perturbation profiles induced by the two different probes, specific interactions of [Gd2(L7)(H2O)2] with the enzyme could be ruled out. The large size of the latter probe is suggested to be responsible for the strong paramagnetic perturbations observed for CalphaH groups which are located in convex surface-exposed regions. The combined use of the two probes reveals fine details of the dynamics controlling their approach toward the protein surface.
Bernini
Andrea
A
Biomolecular Structure Research Center and Dipartimento di Biologia Molecolare, Università di Siena, I-53100 Siena, Italy.
Spiga
Ottavia
O
Venditti
Vincenzo
V
Prischi
Filippo
F
Bracci
Luisa
L
Tong
Angela Pui-Ling
AP
Wong
Wing-Tak
WT
Niccolai
Neri
N
D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D005682:Gadolinium / Q000737:chemistry; D008958:Models, Molecular; D015394:Molecular Structure; D009113:Muramidase / Q000737:chemistry*; D009942:Organometallic Compounds / Q000737:chemistry*
D009942:Organometallic Compounds; D005682:Gadolinium; D009113:Muramidase