|
|
|
|
| LEADER |
01000naa a22002652 4500 |
| 001 |
NLM231658265 |
| 003 |
DE-627 |
| 005 |
20231224091243.0 |
| 007 |
cr uuu---uuuuu |
| 008 |
231224s2013 xx |||||o 00| ||eng c |
| 024 |
7 |
|
|a 10.1107/S0909049513020943
|2 doi
|
| 028 |
5 |
2 |
|a pubmed24n0772.xml
|
| 035 |
|
|
|a (DE-627)NLM231658265
|
| 035 |
|
|
|a (NLM)24121328
|
| 040 |
|
|
|a DE-627
|b ger
|c DE-627
|e rakwb
|
| 041 |
|
|
|a eng
|
| 100 |
1 |
|
|a Nakamura, Akihiko
|e verfasserin
|4 aut
|
| 245 |
1 |
0 |
|a Phase-diagram-guided method for growth of a large crystal of glycoside hydrolase family 45 inverting cellulase suitable for neutron structural analysis
|
| 264 |
|
1 |
|c 2013
|
| 336 |
|
|
|a Text
|b txt
|2 rdacontent
|
| 337 |
|
|
|a ƒaComputermedien
|b c
|2 rdamedia
|
| 338 |
|
|
|a ƒa Online-Ressource
|b cr
|2 rdacarrier
|
| 500 |
|
|
|a Date Completed 22.05.2014
|
| 500 |
|
|
|a Date Revised 21.10.2021
|
| 500 |
|
|
|a published: Print-Electronic
|
| 500 |
|
|
|a Citation Status MEDLINE
|
| 520 |
|
|
|a Neutron protein crystallography (NPC) is a powerful tool for determining the hydrogen position and water orientation in proteins, but a much larger protein crystal is needed for NPC than for X-ray crystallography, and thus crystal preparation is a bottleneck. To obtain large protein crystals, it is necessary to know the properties of the target protein in the crystallization solution. Here, a crystal preparation method of fungal cellulase PcCel45A is reported, guided by the phase diagram. Nucleation and precipitation conditions were determined by sitting-drop vapor diffusion. Saturation and unsaturation conditions were evaluated by monitoring crystal dissolution, and a crystallization phase diagram was obtained. To obtain a large crystal, crystallization solution was prepared on a sitting bridge (diameter = 5 mm). Initial crystallization conditions were 40 µl of crystallization solution (40 mg ml(-1) protein with 30.5% 3-methyl-1,5-pentanediol in 50 mM tris-HCl pH 8.0) with a 1,000 µl reservoir (61% 3-methyl-1,5,-pentanediol in 50 mM tris-HCl pH 8.0) at 293 K. After the first crystal appeared, the concentration of precipitant in the reservoir solution was reduced to 60% to prevent formation of further crystals. Finally, we obtained a crystal of 6 mm(3) volume (3 mm × 2 mm × 1 mm), which was suitable for neutron diffraction
|
| 650 |
|
4 |
|a Journal Article
|
| 650 |
|
4 |
|a Research Support, Non-U.S. Gov't
|
| 650 |
|
4 |
|a cellulase
|
| 650 |
|
4 |
|a crystallization phase diagram
|
| 650 |
|
4 |
|a neutron protein crystallography
|
| 650 |
|
7 |
|a Glycoside Hydrolases
|2 NLM
|
| 650 |
|
7 |
|a EC 3.2.1.-
|2 NLM
|
| 650 |
|
7 |
|a Cellulase
|2 NLM
|
| 650 |
|
7 |
|a EC 3.2.1.4
|2 NLM
|
| 700 |
1 |
|
|a Ishida, Takuya
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Fushinobu, Shinya
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Kusaka, Katsuhiro
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Tanaka, Ichiro
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Inaka, Koji
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Higuchi, Yoshiki
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Masaki, Mika
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Ohta, Kazunori
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Kaneko, Satoshi
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Niimura, Nobuo
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Igarashi, Kiyohiko
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Samajima, Masahiro
|e verfasserin
|4 aut
|
| 773 |
0 |
8 |
|i Enthalten in
|t Journal of synchrotron radiation
|d 1994
|g 20(2013), Pt 6 vom: 07. Nov., Seite 859-63
|w (DE-627)NLM09824129X
|x 1600-5775
|7 nnns
|
| 773 |
1 |
8 |
|g volume:20
|g year:2013
|g number:Pt 6
|g day:07
|g month:11
|g pages:859-63
|
| 856 |
4 |
0 |
|u http://dx.doi.org/10.1107/S0909049513020943
|3 Volltext
|
| 912 |
|
|
|a GBV_USEFLAG_A
|
| 912 |
|
|
|a SYSFLAG_A
|
| 912 |
|
|
|a GBV_NLM
|
| 912 |
|
|
|a GBV_ILN_40
|
| 912 |
|
|
|a GBV_ILN_350
|
| 912 |
|
|
|a GBV_ILN_2005
|
| 951 |
|
|
|a AR
|
| 952 |
|
|
|d 20
|j 2013
|e Pt 6
|b 07
|c 11
|h 859-63
|