Protein crystallization method in gel capsules with nucleants for contactless handling of crystals
| Project/Area Number |
25650026
|
|---|
| Research Category |
Grant-in-Aid for Challenging Exploratory Research
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Structural biochemistry
|
|---|
| Research Institution | Institute of Physical and Chemical Research |
|---|
Principal Investigator |
Sugahara Michihiro 国立研究開発法人理化学研究所, 放射光科学総合研究センター, 特別研究員 (00415192)
|
|---|
| Project Period (FY) |
2013-04-01 – 2016-03-31
|
| Project Status |
Completed (Fiscal Year 2015)
|
| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
|
|---|
| Keywords | タンパク質結晶化 / ゲル / ゼオライト / 金属有機構造体 / ゲルカプセル |
|---|
| Outline of Final Research Achievements |
A simple technique for protein crystallization in ionically cross-linked polysaccharide (alginate and carrageenan) gel capsules has been developed for contactless handling of crystals in X-ray crystallography. This method is designed to reduce mechanical damage to crystals caused by physical contact between crystal and mount tool and by osmotic shock during various manipulations including cryoprotection, heavy-atom derivatization and ligand soaking experiments. In this study, ten test proteins could be successfully crystallized in gel capsules. Two complete diffraction data sets from crystals of proteins in gel capsules were collected at 100 K without removing the crystals, showing that the crystals had low mosaicities. To demonstrate heavy-atom derivatization, lysozyme crystals were successfully derivatized with Pt atoms within alginate gel capsules. These results suggest that gel capsules prevent serious damage to protein crystals during such experiments.
|
Report
(4 results)
Research Products
(10 results)
-
[Journal Article] Oil-free hyaluronic acid matrix for serial femtosecond crystallography2016
Author(s)
Michihiro Sugahara, Changyong Song, Mamoru Suzuki, Tetsuya Masuda, Shigeyuki Inoue, Takanori Nakane, Fumiaki Yumoto, Eriko Nango, Rie Tanaka, Kensuke Tono, Yasumasa Joti, Takashi Kameshima, Takaki Hatsui, Makina Yabashi, Osamu Nureki, Keiji Numata & So Iwata
-
Journal Title
Scientific Reports
Volume: 6
Issue: 1
Pages: 24484-24484
DOI
NAID
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
-
[Journal Article] Native sulfur/chlorine SAD phasing for serial femtosecond crystallography.2015
Author(s)
Nakane T, Song C, Suzuki M, Nango E, Kobayashi J, Masuda T, Inoue S, Mizohata E, Nakatsu T, Tanaka T, Tanaka R, Shimamura T, Tono K, Joti Y, Kameshima T, Hatsui T, Yabashi M, Nureki O, Iwata S, Sugahara M.
-
Journal Title
Acta Crystallogr D Biol Crystallogr.
Volume: 71
Issue: 12
Pages: 2519-25
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
-
-
-
-
-
-
-
-
