Use of layer silicate for protein crystallization: Effects of Micromica and chlorite powders in hanging drops

Takehara, Masahide; Ino, Keita; Takakusagi, Yoichi; Oshikane, Hiroyuki; Nureki, Osamu; Ebina, Takeo; Mizukami, Fujio; Sakaguchi, Kengo

doi:10.1016/j.ab.2007.10.046

Cited by 18 publications

(26 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, unfortunately, the quality of lysozyme crystals formed in the presence of F-Saps tends to be lower than those in the absence of the nucleant. Our results are consistent with the previous data, which showed that the growth speed of thaumatin crystal formed in the presence of layered silicate is inversely related to their quality [20].…”

Section: Resultssupporting

confidence: 93%

“…All the crystals, regardless of the absence/presence of F-Saps, belonged to the space group P4 3 2 1 2. Indeed, we previously reported no change in the space group of thaumatin crystals formed in the presence of layered silicate [20]. In terms of quality, each completeness and average I/Sigma from the crystals formed in the presence of F-Saps was lower than those obtained from the control.…”

Section: Resultsmentioning

confidence: 75%

“…Our previous study has shown that two layered silicates, a K-tetrasilicic fluoromica with a 2∶1 layer comprised of tetrahedral and octahedral units and a chlorite with a 2∶1∶1 layer, act as nucleants for crystallization of model proteins [20]. The fluoromica promoted nucleation in eight out of the ten proteins tested.…”

Section: Introductionmentioning

confidence: 90%

See 2 more Smart Citations

Heterogeneous Nucleation of Protein Crystals on Fluorinated Layered Silicate

et al. 2011

Self Cite

View full text Add to dashboard Cite

Here, we describe an improved system for protein crystallization based on heterogeneous nucleation using fluorinated layered silicate. In addition, we also investigated the mechanism of nucleation on the silicate surface. Crystallization of lysozyme using silicates with different chemical compositions indicated that fluorosilicates promoted nucleation whereas the silicates without fluorine did not. The use of synthesized saponites for lysozyme crystallization confirmed that the substitution of hydroxyl groups contained in the lamellae structure for fluorine atoms is responsible for the nucleation-inducing property of the nucleant. Crystallization of twelve proteins with a wide range of pI values revealed that the nucleation promoting effect of the saponites tended to increase with increased substitution rate. Furthermore, the saponite with the highest fluorine content promoted nucleation in all the test proteins regardless of their overall net charge. Adsorption experiments of proteins on the saponites confirmed that the density of adsorbed molecules increased according to the substitution rate, thereby explaining the heterogeneous nucleation on the silicate surface.

show abstract

Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 75%

See 1 more Smart Citation

Heterogeneous Nucleation of Protein Crystals on Fluorinated Layered Silicate

et al. 2011

Self Cite

View full text Add to dashboard Cite

show abstract

“…The search for suitable materials which can induce and control nucleation of a wide variety of proteins, has been ongoing for over 30 years and a variety of approaches have been used to promote and control nucleation to obtain diffraction quality protein crystals [7][8][9][10][11][12][13][14][15] . These approaches include the effects of epitaxy, surface morphology, disordered porosity, surface roughness and surface chemistry, as well as the use of various crystalline and amorphous miscellaneous material 7,10,11,13,[16][17][18][19][20] . This work has offered little in terms of systematic methodological understanding, and most studies have been highly empirical in their approach.…”

Section: Introductionmentioning

confidence: 99%

Selective Crystallization of Proteins Using Engineered Nanonucleants

Shah

Williams

Heng

2012

Crystal Growth & Design

View full text Add to dashboard Cite

Abstract:This study reports for the first time a detailed experimental investigation of protein crystallization in engineered nano-confined spaces with both controlled pore diameters and narrow pore size distributions. We propose a systematic approach for controlling the nucleation and crystallization of biological macromolecules based on a relationship between the protein radius of gyration (R g ) and specific pore diameter. A series of nano-nucleants with ordered mesopores having narrow pore size distributions were prepared. The templates were tested for proteins ranging in molecular weight from 14kDa to 450kDa. Well formed protein crystals were obtained on only one of the five presented nanonucleants for all protein cases tested, highlighting the unique template selectivity exhibited by these nucleants. In addition, Concanavalin A and Catalase were both crystallized at ∼2 times lower supersaturation levels than previously reported by any known method. Our observations fully support theoretical studies which predict the enhanced thermodynamic stability of proteins in nanoconfined cavities, including specifically the importance of nucleant pore diameter with respect to protein radius of gyration. The nucleants described here could have major industrial applications for downstream separation and purification of biopharmaceuticals, as well as improved opportunities for the crystallization of complex proteins for structural determination.

show abstract

“…Other candidate nucleants followed like zeolites, silicates, charged surfaces, porous materials etc. and have been tested for multiple proteins (Sugahara et al, 2008, Takehara et al, 2008. Previous results showed that horsehair and dried seaweed showed increased hits when added to sparse-matrix crystallization trials.…”

Section: The Role Of Heterogeneous Substrates In the Process Of Protementioning

confidence: 99%