[NMR paper] Backbone dynamics of the cytotoxic ribonuclease alpha-sarcin by 15N NMR relaxation me

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Backbone dynamics of the cytotoxic ribonuclease alpha-sarcin by 15N NMR relaxation methods.

Related Articles Backbone dynamics of the cytotoxic ribonuclease alpha-sarcin by 15N NMR relaxation methods.

J Biomol NMR. 2002 Dec;24(4):301-16

Authors: Pérez-Cañadillas JM, Guenneugues M, Campos-Olivas R, Santoro J, Martínez del Pozo A, Gavilanes JG, Rico M, Bruix M

The cytotoxic ribonuclease alpha-sarcin is a 150-residue protein that inactivates ribosomes by selectively cleaving a single phosphodiester bond in a strictly conserved rRNA loop. In order to gain insights on the molecular basis of its highly specific activity, we have previously determined its solution structure and studied its electrostatics properties. Here, we complement those studies by analysing the backbone dynamics of alpha-sarcin through measurement of longitudinal relaxation rates R1, off resonance rotating frame relaxation rates R1 rho, and the 15N[1H] NOE of the backbone amide 15N nuclei at two different magnetic field strengths (11.7 and 17.6 T). The two sets of relaxation parameters have been analysed in terms of the reduced spectral density mapping formalism, as well as by the model-free approach. alpha-Sarcin behaves as an axial symmetric rotor of the prolate type (D(axially)/D(radially)=1.16 +/- 0.02) which tumbles with a correlation time tau(m) of 7.54 +/- 0.02 ns. The rotational diffusion properties have been also independently evaluated by hydrodynamic calculations and are in good agreement with the experimental results. The analysis of the internal dynamics reveals that alpha-sarcin is composed of a rigid hydrophobic core and some exposed segments which undergo fast (ps to ns) internal motions. Slower motions in the mu s to ms time scale are less abundant and in some cases can be assigned to specific motional processes. All dynamic data are discussed in relation to the role of some particular residues of alpha-sarcin in the process of recognition of its ribosomal target.

PMID: 12522295 [PubMed - indexed for MEDLINE]



Source: PubMed