PDBsum entry 2oqu

Hydrolase

PDB id: 2oqu

Contents

Protein chain

240 a.a. *

Ligands

SO4

Metals

_XE

_CA

Waters ×327

* Residue conservation analysis

PDB id:

2oqu

Name:

Hydrolase

Title:

High pressure cryocooling of capillary sample cryoprotection and diffraction phasing at long wavelengths

Structure:

Elastase-1. Chain: a. Ec: 3.4.21.36

Source:

Sus scrofa. Pig. Organism_taxid: 9823

Resolution:

1.80Å R-factor: 0.180 R-free: 0.226

Authors:

C.U.Kim,Q.Hao,S.M.Gruner

Key ref:

C.U.Kim et al. (2007). High-pressure cryocooling for capillary sample cryoprotection and diffraction phasing at long wavelengths. Acta Crystallogr D Biol Crystallogr, 63, 653-659. PubMed id: 17452791 DOI: 10.1107/S0907444907011924

Date:

01-Feb-07 Release date: 01-May-07

Protein chain

P00772  (CELA1_PIG) -  Chymotrypsin-like elastase family member 1 from Sus scrofa

Seq: 266 a.a.

Struc: 240 a.a.

Enzyme reactions

• Enzyme class: E.C.3.4.21.36 - pancreatic elastase.
• Reaction: Hydrolysis of proteins, including elastin. Preferential cleavage: Ala-|-Xaa.

DOI no: 10.1107/S0907444907011924

Acta Crystallogr D Biol Crystallogr 63:653-659 (2007)

PubMed id: 17452791

High-pressure cryocooling for capillary sample cryoprotection and diffraction phasing at long wavelengths.

C.U.Kim, Q.Hao, S.M.Gruner.

ABSTRACT

Crystal cryocooling is usually employed to reduce radiation damage during X-ray crystallography. Recently, a high-pressure cryocooling method has been developed which results in excellent diffraction-quality crystals without the use of penetrative cryoprotectants. Three new developments of the method are presented here: (i) Xe-He high-pressure cryocooling for Xe SAD phasing, (ii) native sulfur SAD phasing and (iii) successful cryopreservation of crystals in thick-walled capillaries without additional cryoprotectants other than the native mother liquor. These developments may be useful for structural solution of proteins without the need for selenomethionine incorporation and for high-throughput protein crystallography.

Selected figure(s)

Figure 2.
Figure 2 He high-pressure cryocooling and S SAD phasing of thaumatin. (a) Thaumatin crystal in a polycarbonate capillary at 110 K. The entire sample, the crystal and mother liquor in the capillary, was He high-pressure cryocooled without adding penetrative cryoprotectants. This clear sample could not be obtained by conventional (room-pressure) flash-cryocooling when cryoprotectants were not added. (b) Diffraction image of the thaumatin crystal grown in a polycarbonate capillary that was He high-pressure cryocooled at 170 MPa. The diffuse background scatter from the capillary ranges from 4.5 to 5.5 Å. The lack of ice rings on the image confirms that water vitrification was successfully achieved under high pressure. The resolution limit [I/ (I) equal to] 5.0] is approximately 1.9 Å and the crystal mosaicity is 0.34°. The diffraction spots in the enlarged region look compact. (c) Anomalous difference map (5 level) generated with the refined phases. All 17 sulfurs that naturally present in thaumatin are clearly visible. The shape of the electron density at disulfide bonds is dumbbell-shaped, so two-sulfur positions could be easily distinguished. The peak height was over the 10 level (red) for most of the sulfur sites and the peak at Met112 was even visible at the 15 level. (d) F[o] electron-density map (1 level) after S SAD phasing and density modification at 1.9 Å resolution. The final refined model solved by molecular replacement was superimposed for visual map evaluation. The figure of merit is 0.824 and the map correlation coefficient calculated with the final refined 2F[o] - F[c] density map is 0.85 for the main chain and 0.76 for side chains.

The above figure is reprinted by permission from the IUCr: Acta Crystallogr D Biol Crystallogr (2007, 63, 653-659) copyright 2007.

Figure was selected by an automated process.