AGGREGATION MODES OF THE SPIN MONO-LABELED TYLOPEPTIN B AND HEPTAIBIN PEPTAIBIOTICS IN FROZEN SOLUTIONS OF WEAK POLARITY AS STUDIED BY PELDOR SPECTROSCOPY
A.D. Milov1, Y.D. Tsvetkov1, M. De Zotti2, C. Prinzivalli2, B. Biondi2, F. Formaggio2, C. Toniolo2, M. Gobbo2
1Institute of Chemical Kinetics and Combustion, Institutskaiya str., 3, Novosibirsk, 630090, Russian Federation
tsvetkov@kinetics.nsc.ru
2Institute of Biomolecular Chemistry, Padova Unit, CNR, 35131 Padova, Italy
Ключевые слова: PELDOR, dipole-dipole interactions, nitroxide mono-labeled tylopeptin B, heptabin, peptaibiotics
Страницы: 76-87
Подраздел: ЭПР СПЕКТРОСКОПИЯ
Аннотация
The X-band PELDOR spectroscopy was used to investigate the magnetic dipole-dipole interactions in glassy solutions of nitroxide mono-labeled tylopeptin B and heptaibin peptaibiotics at 77 K. Specifically, a study was performed of the tylopeptin B peptides labeled at either position 3, 8 or 13, denoted as Т3, Т8 and Т13, respectively. The heptaibin analogs labeled at either position 2 or 14, denoted as H2 and H14, respectively, were also investigated. It was shown that in frozen glassy peptide solutions in methanol, the spin labels are randomly distributed over the solvent volume. This result points to the absence of specific dipolar interactions between the peptides under these conditions. However, peptide aggregation was detected in weakly polar methanol/toluene environments. To study the properties of the resulting aggregates, we examined the depth of modulation for the PELDOR traces as a function of the concentration of the peptides in solution and the distances between the spin labels in the aggregates. Based on the concentration dependencies, the number of peptide molecules in the aggregates was estimated. We find that this value ranges from 2 to 3, depending on the position of the spin label in the peptide sequence. The combined analysis of the distance spectra and the number of peptide molecules in the aggregates allows us to suggest that dimer formation is the prevailing mode of self-association. In the case of spin-labeled tylopeptin B, the molecules in the dimer are head-to-head oriented. In addition, the distance spectra of the aggregates show that the C-termini of the molecules in the tylopeptin B dimer are more mobile than the N-termini. This phenomenon leads to an increase in the spread of the distances between the nitroxides as the label position is approaching the peptide C-terminus. For heptaibin, we show that two forms of dimerization (head-to-head and head-to-tail) occur. Finally, in addition to dimers, aggregates containing 3 or 4 peptide molecules, which give broad lines in the distance spectra, are seen in solution.
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