4Z8K

Phycocyanin structure from T. elongatus at 2.5-A from XFEL using a viscous delivery medium for serial femtosecond crystallography

Summary for 4Z8K

• Entry DOI: 10.2210/pdb4z8k/pdb
• Descriptor: C-phycocyanin alpha chain, C-phycocyanin beta chain, PHYCOCYANOBILIN, ... (4 entities in total)
• Functional Keywords: phycocyanin, t.elongatus, phycobiliprotein, sfx, xfel, agarose, viscous delivery medium., photosynthesis
• Biological source: Thermosynechococcus elongatus (strain BP-1); More
• Total number of polymer chains: 2
• Total formula weight: 37453.39

Authors

Fromme, R.,Basu, S.,Fromme, P. (deposition date: 2015-04-09, release date: 2015-08-12, Last modification date: 2023-09-27)

Primary citation

Conrad, C.E.,Basu, S.,James, D.,Wang, D.,Schaffer, A.,Roy-Chowdhury, S.,Zatsepin, N.A.,Aquila, A.,Coe, J.,Gati, C.,Hunter, M.S.,Koglin, J.E.,Kupitz, C.,Nelson, G.,Subramanian, G.,White, T.A.,Zhao, Y.,Zook, J.,Boutet, S.,Cherezov, V.,Spence, J.C.,Fromme, R.,Weierstall, U.,Fromme, P.
A novel inert crystal delivery medium for serial femtosecond crystallography.
Iucrj, 2:421-430, 2015

PubMed Abstract: Serial femtosecond crystallography (SFX) has opened a new era in crystallo-graphy by permitting nearly damage-free, room-temperature structure determination of challenging proteins such as membrane proteins. In SFX, femtosecond X-ray free-electron laser pulses produce diffraction snapshots from nanocrystals and microcrystals delivered in a liquid jet, which leads to high protein consumption. A slow-moving stream of agarose has been developed as a new crystal delivery medium for SFX. It has low background scattering, is compatible with both soluble and membrane proteins, and can deliver the protein crystals at a wide range of temperatures down to 4°C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 Å resolution using 300 µg of microcrystals embedded into the agarose medium post-crystallization. The agarose delivery method reduces protein consumption by at least 100-fold and has the potential to be used for a diverse population of proteins, including membrane protein complexes.

PubMed: 26177184
DOI: 10.1107/S2052252515009811

Experimental method

X-RAY DIFFRACTION (2.5 Å)