PDBsum entry 5hqd

Hydrolase

PDB id: 5hqd

Contents

Protein chain

316 a.a.

Metals

_ZN

_CA ×5

Waters ×269

PDB id:

5hqd

Name:

Hydrolase

Title:

Acoustic injectors for drop-on-demand serial femtosecond crystallography

Structure:

Thermolysin. Chain: a. Engineered: yes

Source:

Thermus thermophilus. Organism_taxid: 274. Expressed in: thermus thermophilus. Expression_system_taxid: 274

Resolution:

2.52Å R-factor: 0.222 R-free: 0.288

Authors:

C.G.Roesser,R.Agarwal,M.Allaire,R.Alonso-Mori,B.Andi,J.F.R.Bachega, M.Bommer,A.S.Brewster,M.C.Browne,R.Chatterjee,E.Cho,A.E.Cohen, M.Cowan,S.Datwani,V.L.Davidson,J.Defever,B.Eaton,R.Ellson,Y.Feng, L.P.Ghislain,J.M.Glownia,G.Han,J.Hattne,J.Hellmich,A.Heroux, M.Ibrahim,J.Kern,A.Kuczewski,H.T.Lemke,P.Liu,L.Majlof, W.M.Mcclintock,S.Myers,S.Nelsen,J.Olechno,A.M.Orville,N.K.Sauter, A.S.Soares,M.S.Soltis,H.Song,R.G.Stearns,R.Tran,Y.Tsai, M.Uervirojnangkoorn,C.M.Wilmot,V.Yachandra,J.Yano,E.T.Yukl,D.Zhu, A.Zouni

Key ref:

C.G.Roessler et al. (2016). Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography. Structure, 24, 631-640. PubMed id: 26996959 DOI: 10.1016/j.str.2016.02.007

Date:

21-Jan-16 Release date: 10-Feb-16

Supersedes:

5f80

Protein chain

V5IRV7  (V5IRV7_THETH) -  Neutral metalloproteinase from Thermus thermophilus

Seq: 317 a.a.

Struc: 316 a.a.

Enzyme reactions

• Enzyme class: E.C.3.4.24.- - ?????

DOI no: 10.1016/j.str.2016.02.007

Structure 24:631-640 (2016)

PubMed id: 26996959

Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography.

C.G.Roessler, R.Agarwal, M.Allaire, R.Alonso-Mori, B.Andi, J.F.Bachega, M.Bommer, A.S.Brewster, M.C.Browne, R.Chatterjee, E.Cho, A.E.Cohen, M.Cowan, S.Datwani, V.L.Davidson, J.Defever, B.Eaton, R.Ellson, Y.Feng, L.P.Ghislain, J.M.Glownia, G.Han, J.Hattne, J.Hellmich, A.Héroux, M.Ibrahim, J.Kern, A.Kuczewski, H.T.Lemke, P.Liu, L.Majlof, W.M.McClintock, S.Myers, S.Nelsen, J.Olechno, A.M.Orville, N.K.Sauter, A.S.Soares, S.M.Soltis, H.Song, R.G.Stearns, R.Tran, Y.Tsai, M.Uervirojnangkoorn, C.M.Wilmot, V.Yachandra, J.Yano, E.T.Yukl, D.Zhu, A.Zouni.

ABSTRACT

X-ray free-electron lasers (XFELs) provide very intense X-ray pulses suitable for macromolecular crystallography. Each X-ray pulse typically lasts for tens of femtoseconds and the interval between pulses is many orders of magnitude longer. Here we describe two novel acoustic injection systems that use focused sound waves to eject picoliter to nanoliter crystal-containing droplets out of microplates and into the X-ray pulse from which diffraction data are collected. The on-demand droplet delivery is synchronized to the XFEL pulse scheme, resulting in X-ray pulses intersecting up to 88% of the droplets. We tested several types of samples in a range of crystallization conditions, wherein the overall crystal hit ratio (e.g., fraction of images with observable diffraction patterns) is a function of the microcrystal slurry concentration. We report crystal structures from lysozyme, thermolysin, and stachydrine demethylase (Stc2). Additional samples were screened to demonstrate that these methods can be applied to rare samples.