X-ray Transparent Microfluidic Chip for Mesophase-Based Crystallization of Membrane Proteins and On-Chip Structure Determination

Khvostichenko, Daria S.; Schieferstein, Jeremy M.; Pawate, Ashtamurthy S.; Laible, Philip D.; Kenis, Paul J. A.

doi:10.1021/cg5011488

Title: X-ray Transparent Microfluidic Chip for Mesophase-Based Crystallization of Membrane Proteins and On-Chip Structure Determination

Journal Article · Wed Sep 10 00:00:00 EDT 2014 · Crystal Growth and Design

DOI:https://doi.org/10.1021/cg5011488· OSTI ID:1158727

Khvostichenko, Daria S. ^[1]; Schieferstein, Jeremy M. ^[1]; Pawate, Ashtamurthy S. ^[1]; Laible, Philip D. ^[2]; Kenis, Paul J. A. ^[1]

Department of Chemical & Biomolecular Engineering, University of Illinois at Urbana−Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, United States
Biosciences Division, Argonne National Laboratory, Argonne, Illinois 60439, United States

Crystallization from lipidic mesophase matrices is a promising route to diffraction-quality crystals and structures of membrane proteins. The microfluidic approach reported here eliminates two bottlenecks of the standard mesophase-based crystallization protocols: (i) manual preparation of viscous mesophases and (ii) manual harvesting of often small and fragile protein crystals. In the approach reported here, protein-loaded mesophases are formulated in an X-ray transparent microfluidic chip using only 60 nL of the protein solution per crystallization trial. The X-ray transparency of the chip enables diffraction data collection from multiple crystals residing in microfluidic wells, eliminating the normally required manual harvesting and mounting of individual crystals. In addition, we validated our approach by on-chip crystallization of photosynthetic reaction center, a membrane protein from Rhodobacter sphaeroides, followed by solving its structure to a resolution of 2.5 Å using X-ray diffraction data collected on-chip under ambient conditions. A moderate conformational change in hydrophilic chains of the protein was observed when comparing the on-chip, room temperature structure with known structures for which data were acquired under cryogenic conditions.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Advanced Research Projects Agency - Energy (ARPA-E)

Grant/Contract Number:: AC02-06CH11357; R01 GM086727

OSTI ID:: 1158727

Alternate ID(s):: OSTI ID: 1225366

Journal Information:: Crystal Growth and Design, Journal Name: Crystal Growth and Design Vol. 14 Journal Issue: 10; ISSN 1528-7483

Publisher:: American Chemical SocietyCopyright Statement

Country of Publication:: France

Language:: English

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Cited by: 30 works

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