標題: Spatially Precise, Soft Microseeding of Single Protein Crystals by Femtosecond Laser Ablation
作者: Yoshikawa, Hiroshi Y.
Hosokawa, Yoichiroh
Murai, Ryota
Sazaki, Gen
Kitatani, Tomoya
Adachi, Hiroaki
Inoue, Tsuyoshi
Matsumura, Hiroyoshi
Takano, Kazufumi
Murakami, Satoshi
Nakabayashi, Seiichiro
Mori, Yusuke
Masuhara, Hiroshi
應用化學系
應用化學系分子科學碩博班
Department of Applied Chemistry
Institute of Molecular science
公開日期: 1-九月-2012
摘要: We developed a spatially precise, soft microseeding method for the production of single protein crystals that are suitable for X-ray crystallographic studies. We used focused femtosecond laser pulses to produce, via multiphoton absorption processes, seed crystals from small regions (similar to 1 mu m(2)) of crystals. Hen egg-white lysozyme seed crystals, produced in this manner, grew to be single crystals without any deterioration in their crystallinity. We also validated the technique using polycrystals for the membrane protein, acriflavine resistance protein B, for which single crystals are very difficult to obtain. In addition, we found that the shape of a tetragonal lysozyme crystal prepared from the seed could be controlled by altering the time interval between the initiation of crystallization and laser ablation. We also tried to comprehend the mechanism of femtosecond laser-induced microseeding. We visualized the ablated surfaces of the lysozyme crystals by atomic force microscopy and by laser confocal microscopy combined with differential interference microscopy. The results obtained in this study clearly demonstrate that femtosecond laser ablation of protein crystals is based on a photomechanical process, which ejects crystal fragments with little thermal damage. Femtosecond laser ablation is indeed very promising to produce high quality protein seed crystals from polycrystals or cracked crystals that are not suitable for X-ray diffraction studies.
URI: http://dx.doi.org/10.1021/cg300018t
http://hdl.handle.net/11536/20482
ISSN: 1528-7483
DOI: 10.1021/cg300018t
期刊: CRYSTAL GROWTH & DESIGN
Volume: 12
Issue: 9
起始頁: 4334
結束頁: 4339
顯示於類別:期刊論文


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