Direct observation of adsorption sites of protein impurities and their effects on step advancement of protein crystals

	Direct observation of adsorption sites of protein impurities and their effects on step advancement of protein crystals
	Van Driessehe Alexander ES; Sazaki G; Dai GL(戴国亮); Otalora F; Gavira Ja; Matsui T; Yoshizaki I; Tsukamoto K; Nakajima K
刊名	Crystal Growth & Design
	2009
通讯作者邮箱	sazaki@lowtem.hokudai.ac.jp
卷号	9 期号:7 页码:3062-3071
关键词	Polypeptides Dimers Covalent bonds Fluorescent material Crystal faces Growth mechanism Interference microscopy Confocal microscopy Lysozyme Tetragonal crystals Impurity effect Macromolecules Proteins Adsorption site
ISSN号	1528-7483
通讯作者	Sazaki G
合作状况	国际
中文摘要	We measured noninvasively step velocities of elementary two-dimensional (2D) islands on {110} faces of tetragonal lysozyme crystals, under various supersaturations, by laser confocal microscopy combined with differential interference contrast microscopy. We studied the correlation between the effects of protein impurities on the growth of elementary steps and their adsorption sites on a crystal surface, using three kinds of proteins: fluorescent-labeled lysozyme (F-lysozyme), covalently bonded dimers of lysozyme (dimer), and a 18 kDa polypeptide (18 kDa). These three protein impurities suppressed the advancement of the steps. However, they exhibited different supersaturation dependencies of the suppression of the step velocities. To clarify the cause of this difference, we observed in situ the adsorption sites of individual molecules of F-lysozyme and fluorescent-labeled dimer (F-dimer) on the crystal surface by single-molecule visualization. We found that F-lysozyme adsorbed preferentially on steps (i.e., kinks), whereas F-dimer adsorbed randomly on terraces. Taking into account the different adsorption sites of F-lysozyme and F-dimer, we could successfully explain the different effects of the impurities on the step velocities. These observations strongly suggest that 18 kDa also adsorbs randomly on terraces. Seikagaku lysozyme exhibited a complex effect that could not alone be explained by the two major impurities (dimer and 18 kDa) present in Seikagaku lysozyme, indicating that trace amounts of other impurities significantly affect the step advancement.
学科主题	交叉与边缘领域的力学
类目[WOS]	Chemistry, Multidisciplinary ; Crystallography ; Materials Science, Multidisciplinary
研究领域[WOS]	Chemistry ; Crystallography ; Materials Science
关键词[WOS]	EGG-WHITE LYSOZYME ; REFLECTION FLUORESCENCE MICROSCOPY ; LOCALLY WEIGHTED REGRESSION ; ADVANCED OPTICAL MICROSCOPY ; GROWTH-KINETICS ; MACROMOLECULAR IMPURITIES ; MOLECULAR-MECHANISMS ; DEFECT FORMATION ; CRYSTALLIZATION ; MICROHETEROGENEITY
收录类别	SCI
语种	英语
WOS记录号	WOS:000267609600019
公开日期	2009-12-16
内容类型	期刊论文
源URL	[http://dspace.imech.ac.cn/handle/311007/28715]
专题	力学研究所_国家微重力实验室
推荐引用方式 GB/T 7714	Van Driessehe Alexander ES,Sazaki G,Dai GL,et al. Direct observation of adsorption sites of protein impurities and their effects on step advancement of protein crystals[J]. Crystal Growth & Design,2009,9(7):3062-3071.
APA	Van Driessehe Alexander ES.,Sazaki G.,戴国亮.,Otalora F.,Gavira Ja.,...&Nakajima K.(2009).Direct observation of adsorption sites of protein impurities and their effects on step advancement of protein crystals.Crystal Growth & Design,9(7),3062-3071.
MLA	Van Driessehe Alexander ES,et al."Direct observation of adsorption sites of protein impurities and their effects on step advancement of protein crystals".Crystal Growth & Design 9.7(2009):3062-3071.