Systematic use of the incomplete factorial approach in the design of protein crystallization experiments.
We have used the Incomplete Factorial Approach (Carter, C. W., and Carter, C. W., Jr. (1979) J. Biol. Chem. 254, 12219-12223) in conjunction with the program Cristal (Roussel, A., Serre, L., Frey M., and Fontecilla-Camps, J. (1990) J. Crystal Growth 106, 405-409) to crystallize six different proteins. We were able to obtain crystals and to identify the critical factors for crystallization for each of these six proteins. In some of the cases, we succeeded on the first try while using only minute amounts of protein. This study proves that the Incomplete Factorial Approach is a powerful tool in identifying the factors that need to be varied to achieve crystallization. Single crystals of adequate size were obtained for all the proteins reported here, although some did not diffract well enough to be studied by x-ray diffraction methods; the asymmetric units of these latter crystals contain a large metric units of these latter crystals contain a large number of molecules, which is most likely due to the presence of significant amounts of carbohydrate in the proteins.
Abergel
C
C
Laboratoire de Cristallographie et de Cristallisation des Macromolécules Biologiques, URA 1296-Centre National de la Recherche Scientifique, Faculté de Médecine Secteur-Nord, Marseille, France.
Moulard
M
M
Moreau
H
H
Loret
E
E
Cambillau
C
C
Fontecilla-Camps
J C
JC
D000818:Animals; D002417:Cattle; D003460:Crystallization; D004285:Dogs; D006801:Humans; D008049:Lipase / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D011817:Rabbits; D013270:Stomach / Q000201:enzymology
D011506:Proteins; D008049:Lipase