Processing of membrane protein crystal using ultraviolet laser irradiation.
We demonstrated the processing of a membrane protein crystal, using a pulsed UV laser soft ablation (PULSA) technique. Irradiation with deep-UV laser pulses at a wavelength of 193 nm successfully processed not only single crystals of the membrane transporter protein AcrB but also nylon loops and cryoprotectants at a cryogenic temperature. Nonprocessed parts of the crystals exhibited no signs of crack or denaturation after the laser exposure. The trimmed crystals were found to be of high resolution for X-ray diffraction data collection. The results described here indicate that PULSA processing is an effective tool for membrane protein crystals, as well as for soluble protein crystals.
Kitano
Hiroshi
H
Department of Electrical Engineering, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Murakami
Satoshi
S
Adachi
Hiroaki
H
Matsumura
Hiroyoshi
H
Takano
Kazufumi
K
Inoue
Tsuyoshi
T
Mori
Yusuke
Y
Doi
Masaaki
M
Sasaki
Takatomo
T
D002352:Carrier Proteins / Q000032:analysis / Q000737:chemistry* / Q000528:radiation effects* / Q000648:ultrastructure; D003460:Crystallization / Q000379:methods*; D029968:Escherichia coli Proteins / Q000032:analysis / Q000737:chemistry* / Q000528:radiation effects* / Q000648:ultrastructure; D007834:Lasers; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry* / Q000528:radiation effects* / Q000648:ultrastructure; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D046912:Multiprotein Complexes / Q000032:analysis / Q000737:chemistry / Q000528:radiation effects / Q000648:ultrastructure; D011208:Powders; D014466:Ultraviolet Rays
D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D046912:Multiprotein Complexes; D011208:Powders; C083496:AcrB protein, E coli