Structure and growth of ultrasmall protein microcrystals by synchrotron radiation: II. µGISAX and microscopy of lysozyme

Abstract

The early steps of growth and nucleation of the lysozyme microcrystals by classical and nanotemplate-based hanging vapor diffusion methods are studied using microGISAXS at the European Synchrotron Radiation Facility (ESRF) in Grenoble, France. Out-of-plane cuts in the Yoneda regions of the 2D scattering profiles point to the detection of ultrasmall lysozyme crystals by microGISAXS quite before than by light microscopy. Furthermore lysozyme crystal formation occurs quite earlier with the nanotemplate than with the classical method. Our data are compatible with two distinct modes of crystal nucleation and growth for P450sc and lysozyme.

Structure and growth of ultrasmall protein microcrystals by synchrotron radiation: I. µGISAXS and µdiffraction of P450scc

“…Several methods of interpreting the data are indeed only mentioned, but not implemented because of the extreme difficulty in utilizing ab initio considerations at the present stage of development. The interpretations was stopped there simply in showing images of the Yoneda peak, while here continue in more depth with in‐ and out‐of‐ planes data being presented and discussed in both cytochrome (this study) and lysozyme [accompanying study by Pechkova and Nicolini, 2005].…”

“…In every drop protein solution pH 7.4, 1M NaCl, 0.3 NaColate is mixed in proportion (1:1) with precipitating buffer pH 7.4, 0.1M (NH 4 ) 2 SO 4 , 20% PEG 400. Further details on the parameters of the P450scc microcrystal growth are provided in the accompanying study [Pechkova and Nicolini, 2002a,b, 2005].…”

In situ Monitoring By Raman Spectroscopy of Lysozyme Conformation during “Nanotemplate” Induced Crystallization

“…Raman-based measurements of biological samples have already been exploited for the identification of molecular specific markers for disease detection and monitoring [1]. The use of fiber optic technology coupled with Raman spectroscopy is ideal for application to aqueous solutions, either with or without Langmuir-Blodgett (LB) nanotemplate [2,3]. Raman is expected to evaluate the lysozyme concentration changes in vapour diffusion hanging drop method with and without LangmuirBlodgett nanotemplate.…”

Structural Changes of Casein Micelles in a Calcium Gradient Film

“…Compared with other surface sensitive biophysical probes like AFM21, 11 or electron microscopy,6, 10 GISAXS is a non‐destructive technique with a high sensitivity and statistical relevance. Crystal growth and nucleation induced by highly ordered protein nanotemplates were recently investigated 22–24…”