High pressure-jump apparatus for kinetic studies of protein folding reactions using the small-angle synchrotron x-ray scattering technique

An apparatus suitable for pressure-jump experiments with variable pressure amplitude and a fast response time to facilitate time-resolved small-angle x-ray scattering at synchrotron facilities is described. The high pressure-jump apparatus is capable of performing bidirectional pressure jumps at a time resolution as high as 5 ms. The high pressure sample cell presented has flat diamond windows and is suited for pressures up to 0.7 GPa operating in the temperature range from −40 to 120 °C. The cell is designed for investigating biological and other soft condensed matter materials. Modifications on the window supports allow also simultaneous wide-angle x-ray scattering data to be taken. We have used the equipment to study the kinetics of protein folding reactions. The performance of the apparatus is demonstrated by presenting data on the pressure-induced un/refolding reaction of the water-soluble protein SNase WT.

[1]  R. Templer,et al.  Kinetics of Lyotropic Phase Transitions Involving the Inverse Bicontinuous Cubic Phases , 2000 .

[2]  R. Winter,et al.  Kinetics of phase transformations between lyotropic mesophases of different topology: a time-resolved synchrotron X-ray diffraction study using the pressure-jump relaxation. , 2000 .

[3]  K. Hodgson,et al.  Transient dimer in the refolding kinetics of cytochrome c characterized by small-angle X-ray scattering. , 1999, Biochemistry.

[4]  S. Seifert,et al.  Design and performance of a ASAXS instrument at the Advanced Photon Source , 1999 .

[5]  D. Baker,et al.  Chain collapse can occur concomitantly with the rate-limiting step in protein folding , 1999, Nature Structural Biology.

[6]  H. Amenitsch,et al.  High-pressure instrument for small- and wide-angle x-ray scattering. II. Time-resolved experiments , 1999 .

[7]  J. Jonas,et al.  High Pressure Molecular Science , 1999 .

[8]  J. Jonas.  High-Pressure Raman Scattering Studies of Fluids , 1999 .

[9]  C. Czeslik,et al.  Effect of pressure on the stability, phase behaviour and transformation kinetics between structures of lyotropic lipid mesophases and model membrane systems , 1998 .

[10]  T. Fujisawa,et al.  High-pressure solution X-ray scattering of protein using a hydrostatic cell with diamond windows. , 1998, Journal of synchrotron radiation.

[11]  C. Royer,et al.  Structural characterization of the pressure-denatured state and unfolding/refolding kinetics of staphylococcal nuclease by synchrotron small-angle X-ray scattering and Fourier-transform infrared spectroscopy. , 1998, Journal of molecular biology.

[12]  M. Kriechbaum,et al.  High pressure cell for small- and wide-angle x-ray scattering , 1997 .

[13]  R. Templer,et al.  A PRESSURE- AND TEMPERATURE-CONTROLLED SAMPLE CELL FOR THE STRUCTURAL INVESTIGATION OF LYOTROPIC LIQUID CRYSTALS USING A SMALL-ANGLE X-RAY DIFFRACTION LINE SOURCE , 1996 .

[14]  R. Winter,et al.  Rate of phase transformations between mesophases of the 1:2 lecithin/fatty acid mixtures DMPC/MA and DPPC/PA : a time-resolved synchrotron X-ray diffraction study , 1996 .

[15]  C. Czeslik,et al.  High pressure synchrotron X-ray diffraction studies of biological molecules using the diamond anvil technique , 1996 .

[16]  S. Gruner.  Time-resolved x-ray diffraction of biological materials. , 1987, Science.

[17]  F. Richards,et al.  Collection and processing of X‐ray diffraction data from protein crystals at high pressure , 1986 .

[18]  C. Pistorius,et al.  Phase Diagrams of H2O and D2O at High Pressures , 1968 .