A new manual dispensing system for in meso membrane protein crystallization with using a stepping motor-based dispenser

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• 作者：Masakatsu Hato (1) (2)
  Toshiaki Hosaka (1) (2)
  Hiroaki Tanabe (1) (2)
  Tokuji Kitsunai (3)
  Shigeyuki Yokoyama (1) (4)
  
• 关键词：In meso membrane protein crystallization ; Dispenser ; Manual dispensing ; Cubic phase ; Membrane protein ; Bacteriorhodopsin
• 刊名：Journal of Structural and Functional Genomics
• 出版年：2014
• 出版时间：September 2014
• 年：2014
• 卷：15
• 期：3
• 页码：165-171
• 全文大小：867 KB
• 参考文献：1. Landau EM, Rosenbusch JP (1996) Lipidic cubic phases: a novel concept for the crystallization of membrane proteins. Proc Natl Acad Sci USA 93:14532-4535 CrossRef
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  9. Hato M, Minamikawa H, Salkar RA, Matsutani S (2002) Alkylglycosides with an isoprenoid-type hydrophobic chain can afford greater control of aqueous phase structures at low temperatures. Langmuir 18:3425-429 CrossRef
  10. Hato M, Yamashita I, Kato T, Abe Y (2004) Aqueous phase behavior of a 1-O-phytanyl-β-D-xyloside/water system. glycolipid-based bicontinuous cubic phases of crystallographic space groups / Pn3m and / Ia3d. Langmuir 20:11366-1373 CrossRef
  11. Hato M, Yamashita J, Shiono M (2009) Aqueous phase behavior of lipids with isoprenoid type hydrophobic chains. J Phys Chem B 113:10196-0209
  12. Yamashita J, Shiono M, Hato M (2008) New lipid family that forms inverted cubic phases in equilibrium with excess water: molecular structure-aqueous phase structure relationship for lipids with 5, 9, 13, 17-tetramethyloctadecyl and 5, 9, 13, 17-tetramethyloctadecanoyl chains. J Phys Chem B 112:12286-2296
  
• 作者单位：Masakatsu Hato (1) (2)
  Toshiaki Hosaka (1) (2)
  Hiroaki Tanabe (1) (2)
  Tokuji Kitsunai (3)
  Shigeyuki Yokoyama (1) (4)
  
  1. RIKEN Systems and Structural Biology Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan
  2. Division of Structural and Synthetic Biology, RIKEN Center for Life Science Technologies, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan
  3. Advanced Photonics Technology Development Group, RIKEN Center for Advanced Photonics, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan
  4. RIKEN Structural Biology Laboratory, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan
  
• ISSN：1570-0267

文摘

A reliable and easy to use manual dispensing system has been developed for the in meso membrane protein crystallization method. The system consists of a stepping motor-based dispenser with a new microsyringe system for dispensing, which allows us to deliver any desired volume of highly viscous lipidic mesophase in the range from ~50 to at least ~200?nl. The average, standard deviation, and coefficient of variation of 20 repeated deliveries of 50?nl cubic phase were comparable to those of a current robotic dispensing. Moreover, the bottom faces of boluses delivered to the glass crystallization plate were reproducibly circular in shape, and their centers were within about 100?μm from the center of the crystallization well. The system was useful for crystallizing membrane and soluble proteins in meso.