|In situ observation of elementary growth processes of protein crystals by advanced optical microscopy|
|Sazaki G; Van Driessche AES; Dai GL(戴国亮); Okada M; Matsui T; Otalora F; Tsukamoto K; Nakajima K; Sazaki, G; Hokkaido Univ, Inst Low Temp Sci, Kita Ku, N19-W8, Sapporo, Hokkaido 0600819, Japan.
|Source Publication||PROTEIN AND PEPTIDE LETTERS
|Abstract||To start systematically investigating the quality improvement of protein crystals, the elementary growth processes of protein crystals must be first clarified comprehensively. Atomic force microscopy (AFM) has made a tremendous contribution toward elucidating the elementary growth processes of protein crystals and has confirmed that protein crystals grow layer by layer utilizing kinks on steps, as in the case of inorganic and low-molecular-weight compound crystals. However, the scanning of the AFM cantilever greatly disturbs the concentration distribution and solution flow in the vicinity of growing protein crystals. AFM also cannot visualize the dynamic behavior of mobile solute and impurity molecules on protein crystal surfaces. To compensate for these disadvantages of AFM, in situ observation by two types of advanced optical microscopy has been recently performed. To observe the elementary steps of protein crystals noninvasively, laser confocal microscopy combined with differential interference contrast microscopy (LCM-DIM) was developed. To visualize individual mobile protein molecules, total internal reflection fluorescent (TIRF) microscopy, which is widely used in the field of biological physics, was applied to the visualization of protein crystal surfaces. In this review, recent progress in the noninvasive in situ observation of elementary steps and individual mobile protein molecules on protein crystal surfaces is outlined.|
Advanced Optical Microscopy
In Situ Observation
Single Molecule Visualization
Tetragonal Lysozyme Crystals
Reflection Fluorescence Microscopy
Locally Weighted Regression
Facet Morphology Response
|Funding Organization||The authors thank Y. Saito and S. Kobayashi (Olympus Engineering Co., Ltd.) for their technical support in the development of LCM-DIM, and H. Higuchi (The University of Tokyo) and T. Watanabe (Osaka University) for their technical assistance and valuable discussion on single-molecule visualization experiments. This work was partially supported by Grants-in-Aid Nos. 16360001, 17034007, and 18360003 for Scientific Research from the Ministry of Education, Science and Culture of Japan (G. S.) and Project Research B at the Center for Interdisciplinary Research, Tohoku University (G.S. and G.D.), Grant No. AYA2009-10655 from the Ministry of Science and Innovation, Spain (F.O, and A.V.D), and the Consolider-Ingenio 2010 project "Factoria Espanola de cristalizacion" (F.O, and A.V.D).
|Corresponding Author||Sazaki, G; Hokkaido Univ, Inst Low Temp Sci, Kita Ku, N19-W8, Sapporo, Hokkaido 0600819, Japan.|
Sazaki G,Van Driessche AES,Dai GL,et al. In situ observation of elementary growth processes of protein crystals by advanced optical microscopy[J]. PROTEIN AND PEPTIDE LETTERS,2012,19(7):743-760.
Sazaki G.,Van Driessche AES.,Dai GL.,Okada M.,Matsui T.,...&Hokkaido Univ, Inst Low Temp Sci, Kita Ku, N19-W8, Sapporo, Hokkaido 0600819, Japan..(2012).In situ observation of elementary growth processes of protein crystals by advanced optical microscopy.PROTEIN AND PEPTIDE LETTERS,19(7),743-760.
Sazaki G,et al."In situ observation of elementary growth processes of protein crystals by advanced optical microscopy".PROTEIN AND PEPTIDE LETTERS 19.7(2012):743-760.