Protein nanocrystallography, a new technology for crystal growth based on protein nanotemplates, has recently been shown to produce diffracting, stable and radiation-resistant lysozyme crystals. This article, by computing these lysozyme crystals' atomic structures, obtained by the diffraction patterns of microfocused synchrotron radiation, provides a possible mechanism for this increased stability, namely a significant decrease in water content accompanied by a minor but significant α-helix increase. These data are shown to be compatible with the circular dichroism and two-dimensional Fourier transform spectra of high-resolution H NMR of proteins dissolved from the same nanotemplate-based crystal versus those from a classical crystal. Finally, evidence for protein direct transfer from the nanotemplate to the drop and the participation of the template proteins in crystal nucleation and growth is provided by high-resolution NMR spectrometry and mass spectrometry. Furthermore, the lysozyme nanotemplate appears stable up to 523 K, as confirmed by a thermal denaturation study using spectropolarimetry. The overall data suggest that heat-proof lysozyme presence in the crystal provides a possible explanation of the crystal's resistance to synchrotron radiation.
|Titolo:||Protein nanocrystallography : Growth mechanism and atomic structure of crystals induced by nanotemplates|
VASILE, FRANCESCA (Secondo)
|Parole Chiave:||CD; Mass spectroscopy; NMR; Protein crystals; Animals; Chickens; Crystallization; Magnetic Resonance Spectroscopy; Muramidase; Nanotechnology; Mass Spectrometry; Physics and Astronomy (miscellaneous); Instrumentation; Atomic and Molecular Physics, and Optics; Radiation|
|Settore Scientifico Disciplinare:||Settore BIO/11 - Biologia Molecolare|
|Data di pubblicazione:||nov-2005|
|Digital Object Identifier (DOI):||10.1107/S0909049505011647|
|Appare nelle tipologie:||01 - Articolo su periodico|