Monitoring and scoring counter-diffusion protein crystallization experiments in capillaries by in situ dynamic light scattering.
In this paper, we demonstrate the feasibility of using in situ Dynamic Light Scattering (DLS) to monitor counter-diffusion crystallization experiments in capillaries. Firstly, we have validated the quality of the DLS signal in thin capillaries, which is comparable to that obtained in standard quartz cuvettes. Then, we have carried out DLS measurements of a counter-diffusion crystallization experiment of glucose isomerase in capillaries of different diameters (0.1, 0.2 and 0.3 mm) in order to follow the temporal evolution of protein supersaturation. Finally, we have compared DLS data with optical recordings of the progression of the crystallization front and with a simulation model of counter-diffusion in 1D.
Institute for Biochemistry and Molecular Biology, Laboratory for Structural Biology of infection and inflammation, University of Hamburg, Hamburg, Germany.
Dominik
D
Oberthuer
Emilio
E
Melero-García
Karsten
K
Dierks
Arne
A
Meyer
Christian
C
Betzel
Alfonso
A
Garcia-Caballero
Jose A
JA
Gavira
D019747:Aldose-Ketose Isomerases / Q000737:chemistry*; D060166:Capillary Tubing; D003198:Computer Simulation; D003460:Crystallization / Q000295:instrumentation* / Q000379:methods*; D004058:Diffusion / Q000528:radiation effects; D008027:Light; D010316:Particle Size; D011791:Quartz; D012542:Scattering, Radiation; D013302:Streptomyces / Q000201:enzymology; D013997:Time Factors
D011791:Quartz; D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase