Tracing transport of protein aggregates in microgravity versus unit gravity crystallization

Arayik Martirosyan; Sven Falke; Deborah McCombs; Martin Cox; Christopher D. Radka; Jan Knop; Christian Betzel; Lawrence J. DeLucas

doi:10.1038/s41526-022-00191-x

Tracing transport of protein aggregates in microgravity versus unit gravity crystallization

Arayik Martirosyan, Sven Falke, Deborah McCombs, Martin Cox, Christopher D. Radka, Jan Knop, Christian Betzel, Lawrence J. DeLucas

Microbiology, Immunology, and Molecular Genetics

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

Microgravity conditions have been used to improve protein crystallization from the early 1980s using advanced crystallization apparatuses and methods. Early microgravity crystallization experiments confirmed that minimal convection and a sedimentation-free environment is beneficial for growth of crystals with higher internal order and in some cases, larger volume. It was however realized that crystal growth in microgravity requires additional time due to slower growth rates. The progress in space research via the International Space Station (ISS) provides a laboratory-like environment to perform convection-free crystallization experiments for an extended time. To obtain detailed insights in macromolecular transport phenomena under microgravity and the assumed reduction of unfavorable impurity incorporation in growing crystals, microgravity and unit gravity control experiments for three different proteins were designed. To determine the quantity of impurity incorporated into crystals, fluorescence-tagged aggregates of the proteins (acting as impurities) were prepared. The recorded fluorescence intensities of the respective crystals reveal reduction in the incorporation of aggregates under microgravity for different aggregate quantities. The experiments and data obtained, provide insights about macromolecular transport in relation to molecular weight of the target proteins, as well as information about associated diffusion behavior and crystal lattice formation. Results suggest one explanation why microgravity-grown protein crystals often exhibit higher quality. Furthermore, results from these experiments can be used to predict which proteins may benefit more from microgravity crystallization.

Original language	English
Article number	4
Journal	npj Microgravity
Volume	8
Issue number	1
DOIs	https://doi.org/10.1038/s41526-022-00191-x
State	Published - Dec 2022

Bibliographical note

Funding Information:
The authors would like to thank all the people that have been involved in the SpaceX SPX10 and SPX15 mission. The authors would like to thank all the crew members of ISS expedition 50 and 52. Also the authors would like to thank all the participants (i.a. Zin Technologies) of the projects Biophysics 1 and 4. This research has been supported by the NASA (Grant No. 80NSSC18K0013), by Deutsche Luft und Raumfahrt Agentur (DLR) supporting the project via grant 50WB1422, by the Cluster of Excellence ‘Advanced Imaging of Matter’ of the Deutsche Forschungsgemeinschaft (DFG)—EXC 2056—project ID 390715994, and by DFG project BE1443/29-1. We also acknowledge UAB-High Resolution Imaging Service Center, Shelby 135c, Confocal/Light Microscopy Core at the University of Alabama at Birmingham. We acknowledge access to beamlines P11 (DESY) and P13 (DESY/EMBL, Hamburg) and are thankful for technical user support.

Funding Information:
The authors would like to thank all the people that have been involved in the SpaceX SPX10 and SPX15 mission. The authors would like to thank all the crew members of ISS expedition 50 and 52. Also the authors would like to thank all the participants (i.a. Zin Technologies) of the projects Biophysics 1 and 4. This research has been supported by the NASA (Grant No. 80NSSC18K0013), by Deutsche Luft und Raumfahrt Agentur (DLR) supporting the project via grant 50WB1422, by the Cluster of Excellence ?Advanced Imaging of Matter? of the Deutsche Forschungsgemeinschaft (DFG)?EXC 2056?project ID 390715994, and by DFG project BE1443/29-1. We also acknowledge UAB-High Resolution Imaging Service Center, Shelby 135c, Confocal/Light Microscopy Core at the University of Alabama at Birmingham. We acknowledge access to beamlines P11 (DESY) and P13 (DESY/EMBL, Hamburg) and are thankful for technical user support.

Publisher Copyright:
© 2022, The Author(s).

ASJC Scopus subject areas

Medicine (miscellaneous)
Materials Science (miscellaneous)
Biochemistry, Genetics and Molecular Biology (miscellaneous)
Agricultural and Biological Sciences (miscellaneous)
Physics and Astronomy (miscellaneous)
Space and Planetary Science

Access to Document

10.1038/s41526-022-00191-x

Cite this

@article{44534acd34cf45f2b136028b5540d0bf,

title = "Tracing transport of protein aggregates in microgravity versus unit gravity crystallization",

abstract = "Microgravity conditions have been used to improve protein crystallization from the early 1980s using advanced crystallization apparatuses and methods. Early microgravity crystallization experiments confirmed that minimal convection and a sedimentation-free environment is beneficial for growth of crystals with higher internal order and in some cases, larger volume. It was however realized that crystal growth in microgravity requires additional time due to slower growth rates. The progress in space research via the International Space Station (ISS) provides a laboratory-like environment to perform convection-free crystallization experiments for an extended time. To obtain detailed insights in macromolecular transport phenomena under microgravity and the assumed reduction of unfavorable impurity incorporation in growing crystals, microgravity and unit gravity control experiments for three different proteins were designed. To determine the quantity of impurity incorporated into crystals, fluorescence-tagged aggregates of the proteins (acting as impurities) were prepared. The recorded fluorescence intensities of the respective crystals reveal reduction in the incorporation of aggregates under microgravity for different aggregate quantities. The experiments and data obtained, provide insights about macromolecular transport in relation to molecular weight of the target proteins, as well as information about associated diffusion behavior and crystal lattice formation. Results suggest one explanation why microgravity-grown protein crystals often exhibit higher quality. Furthermore, results from these experiments can be used to predict which proteins may benefit more from microgravity crystallization.",

author = "Arayik Martirosyan and Sven Falke and Deborah McCombs and Martin Cox and Radka, {Christopher D.} and Jan Knop and Christian Betzel and DeLucas, {Lawrence J.}",

note = "Funding Information: The authors would like to thank all the people that have been involved in the SpaceX SPX10 and SPX15 mission. The authors would like to thank all the crew members of ISS expedition 50 and 52. Also the authors would like to thank all the participants (i.a. Zin Technologies) of the projects Biophysics 1 and 4. This research has been supported by the NASA (Grant No. 80NSSC18K0013), by Deutsche Luft und Raumfahrt Agentur (DLR) supporting the project via grant 50WB1422, by the Cluster of Excellence {\textquoteleft}Advanced Imaging of Matter{\textquoteright} of the Deutsche Forschungsgemeinschaft (DFG)—EXC 2056—project ID 390715994, and by DFG project BE1443/29-1. We also acknowledge UAB-High Resolution Imaging Service Center, Shelby 135c, Confocal/Light Microscopy Core at the University of Alabama at Birmingham. We acknowledge access to beamlines P11 (DESY) and P13 (DESY/EMBL, Hamburg) and are thankful for technical user support. Funding Information: The authors would like to thank all the people that have been involved in the SpaceX SPX10 and SPX15 mission. The authors would like to thank all the crew members of ISS expedition 50 and 52. Also the authors would like to thank all the participants (i.a. Zin Technologies) of the projects Biophysics 1 and 4. This research has been supported by the NASA (Grant No. 80NSSC18K0013), by Deutsche Luft und Raumfahrt Agentur (DLR) supporting the project via grant 50WB1422, by the Cluster of Excellence ?Advanced Imaging of Matter? of the Deutsche Forschungsgemeinschaft (DFG)?EXC 2056?project ID 390715994, and by DFG project BE1443/29-1. We also acknowledge UAB-High Resolution Imaging Service Center, Shelby 135c, Confocal/Light Microscopy Core at the University of Alabama at Birmingham. We acknowledge access to beamlines P11 (DESY) and P13 (DESY/EMBL, Hamburg) and are thankful for technical user support. Publisher Copyright: {\textcopyright} 2022, The Author(s).",

year = "2022",

month = dec,

doi = "10.1038/s41526-022-00191-x",

language = "English",

volume = "8",

journal = "npj Microgravity",

issn = "2373-8065",

publisher = "Nature Publishing Group",

number = "1",

}

TY - JOUR

T1 - Tracing transport of protein aggregates in microgravity versus unit gravity crystallization

AU - Martirosyan, Arayik

AU - Falke, Sven

AU - McCombs, Deborah

AU - Cox, Martin

AU - Radka, Christopher D.

AU - Knop, Jan

AU - Betzel, Christian

AU - DeLucas, Lawrence J.

N1 - Funding Information: The authors would like to thank all the people that have been involved in the SpaceX SPX10 and SPX15 mission. The authors would like to thank all the crew members of ISS expedition 50 and 52. Also the authors would like to thank all the participants (i.a. Zin Technologies) of the projects Biophysics 1 and 4. This research has been supported by the NASA (Grant No. 80NSSC18K0013), by Deutsche Luft und Raumfahrt Agentur (DLR) supporting the project via grant 50WB1422, by the Cluster of Excellence ‘Advanced Imaging of Matter’ of the Deutsche Forschungsgemeinschaft (DFG)—EXC 2056—project ID 390715994, and by DFG project BE1443/29-1. We also acknowledge UAB-High Resolution Imaging Service Center, Shelby 135c, Confocal/Light Microscopy Core at the University of Alabama at Birmingham. We acknowledge access to beamlines P11 (DESY) and P13 (DESY/EMBL, Hamburg) and are thankful for technical user support. Funding Information: The authors would like to thank all the people that have been involved in the SpaceX SPX10 and SPX15 mission. The authors would like to thank all the crew members of ISS expedition 50 and 52. Also the authors would like to thank all the participants (i.a. Zin Technologies) of the projects Biophysics 1 and 4. This research has been supported by the NASA (Grant No. 80NSSC18K0013), by Deutsche Luft und Raumfahrt Agentur (DLR) supporting the project via grant 50WB1422, by the Cluster of Excellence ?Advanced Imaging of Matter? of the Deutsche Forschungsgemeinschaft (DFG)?EXC 2056?project ID 390715994, and by DFG project BE1443/29-1. We also acknowledge UAB-High Resolution Imaging Service Center, Shelby 135c, Confocal/Light Microscopy Core at the University of Alabama at Birmingham. We acknowledge access to beamlines P11 (DESY) and P13 (DESY/EMBL, Hamburg) and are thankful for technical user support. Publisher Copyright: © 2022, The Author(s).

PY - 2022/12

Y1 - 2022/12

N2 - Microgravity conditions have been used to improve protein crystallization from the early 1980s using advanced crystallization apparatuses and methods. Early microgravity crystallization experiments confirmed that minimal convection and a sedimentation-free environment is beneficial for growth of crystals with higher internal order and in some cases, larger volume. It was however realized that crystal growth in microgravity requires additional time due to slower growth rates. The progress in space research via the International Space Station (ISS) provides a laboratory-like environment to perform convection-free crystallization experiments for an extended time. To obtain detailed insights in macromolecular transport phenomena under microgravity and the assumed reduction of unfavorable impurity incorporation in growing crystals, microgravity and unit gravity control experiments for three different proteins were designed. To determine the quantity of impurity incorporated into crystals, fluorescence-tagged aggregates of the proteins (acting as impurities) were prepared. The recorded fluorescence intensities of the respective crystals reveal reduction in the incorporation of aggregates under microgravity for different aggregate quantities. The experiments and data obtained, provide insights about macromolecular transport in relation to molecular weight of the target proteins, as well as information about associated diffusion behavior and crystal lattice formation. Results suggest one explanation why microgravity-grown protein crystals often exhibit higher quality. Furthermore, results from these experiments can be used to predict which proteins may benefit more from microgravity crystallization.

AB - Microgravity conditions have been used to improve protein crystallization from the early 1980s using advanced crystallization apparatuses and methods. Early microgravity crystallization experiments confirmed that minimal convection and a sedimentation-free environment is beneficial for growth of crystals with higher internal order and in some cases, larger volume. It was however realized that crystal growth in microgravity requires additional time due to slower growth rates. The progress in space research via the International Space Station (ISS) provides a laboratory-like environment to perform convection-free crystallization experiments for an extended time. To obtain detailed insights in macromolecular transport phenomena under microgravity and the assumed reduction of unfavorable impurity incorporation in growing crystals, microgravity and unit gravity control experiments for three different proteins were designed. To determine the quantity of impurity incorporated into crystals, fluorescence-tagged aggregates of the proteins (acting as impurities) were prepared. The recorded fluorescence intensities of the respective crystals reveal reduction in the incorporation of aggregates under microgravity for different aggregate quantities. The experiments and data obtained, provide insights about macromolecular transport in relation to molecular weight of the target proteins, as well as information about associated diffusion behavior and crystal lattice formation. Results suggest one explanation why microgravity-grown protein crystals often exhibit higher quality. Furthermore, results from these experiments can be used to predict which proteins may benefit more from microgravity crystallization.

UR - http://www.scopus.com/inward/record.url?scp=85125336964&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85125336964&partnerID=8YFLogxK

U2 - 10.1038/s41526-022-00191-x

DO - 10.1038/s41526-022-00191-x

M3 - Article

AN - SCOPUS:85125336964

SN - 2373-8065

VL - 8

JO - npj Microgravity

JF - npj Microgravity

IS - 1

M1 - 4

ER -

Tracing transport of protein aggregates in microgravity versus unit gravity crystallization

Abstract

Bibliographical note

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this