TY - JOUR
T1 - Spatially precise, soft microseeding of single protein crystals by femtosecond laser ablation
AU - Yoshikawa, Hiroshi Y.
AU - Hosokawa, Yoichiroh
AU - Murai, Ryota
AU - Sazaki, Gen
AU - Kitatani, Tomoya
AU - Adachi, Hiroaki
AU - Inoue, Tsuyoshi
AU - Matsumura, Hiroyoshi
AU - Takano, Kazufumi
AU - Murakami, Satoshi
AU - Nakabayashi, Seiichiro
AU - Mori, Yusuke
AU - Masuhara, Hiroshi
PY - 2012/9/5
Y1 - 2012/9/5
N2 - 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 (∼1 μm 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.
AB - 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 (∼1 μm 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.
UR - http://www.scopus.com/inward/record.url?scp=84865859220&partnerID=8YFLogxK
U2 - 10.1021/cg300018t
DO - 10.1021/cg300018t
M3 - Article
AN - SCOPUS:84865859220
VL - 12
SP - 4334
EP - 4339
JO - Crystal Growth and Design
JF - Crystal Growth and Design
SN - 1528-7483
IS - 9
ER -