A novel approach for the crystallization of soluble proteins using non-ionic surfactants.
Crystallization trials using three polyoxyethylene surfactants as precipitating agents are described. Of the eight soluble proteins screened, five were successfully crystallized at the first attempt. These included lysozyme, catalase, ferritin, ribonuclease A and ubiquitin. Further work suggested that these surfactants could also be suitable for cryo-crystallographic analysis of crystals. At the concentrations used in the crystallization trials [10-40%(v/v)], they are capable of promoting the formation of non-crystalline glasses at cryogenic temperatures (77K). This would facilitate crystal mounting and allow the minimization of crystal irradiation damage. Results from this study also suggest that proteins remain stable at high concentrations of these surfactants [40%(w/v)] and over long time periods (>1 month). A number of membrane proteins were also screened for crystallization. These included photosystems I and II and light harvesting complexes I and II from spinach and bacteriorhodopsin from Halobacterium halobium++. The trial s were unsuccessful both in the absence and presence of heptane-1,2,3-triol and over a wide range of surfactant concentrations.
Mustafa
A O
AO
Department of Biochemistry and Applied Molecular Biology, UMIST, PO Box88, Manchester M60 1QD, England.
Derrick
J P
JP
Tiddy
G J
GJ
Ford
R C
RC
D002374:Catalase / Q000737:chemistry; D003460:Crystallization; D005293:Ferritins / Q000737:chemistry; D007477:Ions; D008565:Membrane Proteins / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D012995:Solubility; D013501:Surface-Active Agents; D014452:Ubiquitins / Q000737:chemistry
D007477:Ions; D008565:Membrane Proteins; D011506:Proteins; D013501:Surface-Active Agents; D014452:Ubiquitins; D005293:Ferritins; D002374:Catalase; D012259:Ribonuclease, Pancreatic; D009113:Muramidase