Use of layer silicate for protein crystallization: effects of Micromica and chlorite powders in hanging drops.
Two kinds of layer silicate powder, Micromica and chlorite, were used to aid protein crystallization by the addition to hanging drops. Using appropriate crystallization buffers, Micromica powder facilitated crystal growth speed for most proteins tested in this study. Furthermore, the addition of Micromica powder to hanging drops allowed the successful crystallization of lysozyme, catalase, concanavalin A, and trypsin even at low protein concentrations and under buffer conditions that otherwise would not generate protein crystals. Except for threonine synthase and apoferritin, the presence of chlorite delayed crystallization but induced the formation of large crystals. X-ray analysis of thaumatin crystals generated by our novel procedure gave better quality data than did that of crystals obtained by a conventional hanging drop method. Our results suggest that the speed of crystal growth and the quality of the corresponding X-ray data may be inversely related, at least for the formation of thaumatin crystals. The effect of Micromica and chlorite powders and the application of layer silicate powder for protein crystallization are discussed.
Takehara
Masahide
M
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Chiba 278-8510, Japan.
Ino
Keita
K
Takakusagi
Yoichi
Y
Oshikane
Hiroyuki
H
Nureki
Osamu
O
Ebina
Takeo
T
Mizukami
Fujio
F
Sakaguchi
Kengo
K
D000538:Aluminum Silicates; D019757:Carbon-Oxygen Lyases / Q000737:chemistry; D002374:Catalase / Q000737:chemistry; D003208:Concanavalin A / Q000737:chemistry; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray; D009113:Muramidase / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry; D011208:Powders; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D017640:Silicates; D014357:Trypsin / Q000737:chemistry
D000538:Aluminum Silicates; D010940:Plant Proteins; D011208:Powders; D011506:Proteins; D017640:Silicates; D003208:Concanavalin A; C003427:thaumatin protein, plant; D002374:Catalase; D009113:Muramidase; D014357:Trypsin; D019757:Carbon-Oxygen Lyases; C023160:threonine synthase; C011934:mica