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Towards time-resolved serial crystallography in a microfluidic device.

ABSTRACT: Serial methods for crystallography have the potential to enable dynamic structural studies of protein targets that have been resistant to single-crystal strategies. The use of serial data-collection strategies can circumvent challenges associated with radiation damage and repeated reaction initiation. This work utilizes a microfluidic crystallization platform for the serial time-resolved Laue diffraction analysis of macroscopic crystals of photoactive yellow protein (PYP). Reaction initiation was achieved via pulsed laser illumination, and the resultant electron-density difference maps clearly depict the expected pR(1)/pR(E46Q) and pR(2)/pR(CW) states at 10 µs and the pB1 intermediate at 1 ms. The strategies presented here have tremendous potential for extension to chemical triggering methods for reaction initiation and for extension to dynamic, multivariable analyses.

SUBMITTER: Pawate AS 

PROVIDER: S-EPMC4498702 | biostudies-literature | 2015 Jul

REPOSITORIES: biostudies-literature

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Towards time-resolved serial crystallography in a microfluidic device.

Pawate Ashtamurthy S AS   Šrajer Vukica V   Schieferstein Jeremy J   Guha Sudipto S   Henning Robert R   Kosheleva Irina I   Schmidt Marius M   Ren Zhong Z   Kenis Paul J A PJ   Perry Sarah L SL  

Acta crystallographica. Section F, Structural biology communications 20150627 Pt 7

Serial methods for crystallography have the potential to enable dynamic structural studies of protein targets that have been resistant to single-crystal strategies. The use of serial data-collection strategies can circumvent challenges associated with radiation damage and repeated reaction initiation. This work utilizes a microfluidic crystallization platform for the serial time-resolved Laue diffraction analysis of macroscopic crystals of photoactive yellow protein (PYP). Reaction initiation wa  ...[more]

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