| 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
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| Source Publication | Crystal Growth & Design
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| 2009 | |
| Volume | 9Issue:7Pages:3062-3071 |
| ISSN | 1528-7483 |
| Abstract | 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. |
| Keyword | Polypeptides Dimers Covalent Bonds Fluorescent Material Crystal Faces Growth Mechanism Interference Microscopy Confocal Microscopy Lysozyme Tetragonal Crystals Impurity Effect Macromolecules Proteins Adsorption Site |
| Subject Area | 交叉与边缘领域的力学 |
| DOI | 10.1021/cg8006684 |
| Indexed By | SCI |
| Language | 英语 |
| WOS ID | WOS:000267609600019 |
| WOS Keyword | EGG-WHITE LYSOZYME ; REFLECTION FLUORESCENCE MICROSCOPY ; LOCALLY WEIGHTED REGRESSION ; ADVANCED OPTICAL MICROSCOPY ; GROWTH-KINETICS ; MACROMOLECULAR IMPURITIES ; MOLECULAR-MECHANISMS ; DEFECT FORMATION ; CRYSTALLIZATION ; MICROHETEROGENEITY |
| WOS Research Area | Chemistry ; Crystallography ; Materials Science |
| WOS Subject | Chemistry, Multidisciplinary ; Crystallography ; Materials Science, Multidisciplinary |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://dspace.imech.ac.cn/handle/311007/28715 |
| Collection | 微重力重点实验室 |
| Corresponding Author | Sazaki G |
| Recommended Citation 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. |
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