---
OA_place: publisher
OA_type: hybrid
PlanS_conform: '1'
_id: '20258'
abstract:
- lang: eng
  text: The specific introduction of ^1H-^13C or ^1H-^15N moieties into otherwise
    deuterated proteins holds great potential for high-resolution solution and magic-angle
    spinning (MAS) NMR studies of protein structure and dynamics. Arginine residues
    play key roles for example at active sites of enzymes. Taking advantage of a chemically
    synthesized Arg with a ^13C-^1H2 group in an otherwise deuterated backbone, we
    demonstrate here the usefulness of proton-detected MAS NMR approaches to probe
    arginine dynamics. In experiments with crystalline ubiquitin and the 134 kDa tetrameric
    enzyme malate dehydrogenase we detected a wide range of motions, from sites that
    are rigid on time scales of at least tens of milliseconds to residues undergoing
    predominantly nanosecond motions. Spin-relaxation and dipolar-coupling measurements
    enabled quantitative determination of these dynamics. We observed microsecond
    dynamics of residue Arg54 in crystalline ubiquitin, whose backbone is known to
    sample different β-turn conformations on this time scale. The labeling scheme
    and experiments presented here expand the toolkit for high-resolution proton-detected
    MAS NMR.
acknowledged_ssus:
- _id: NMR
- _id: LifeSc
acknowledgement: This work was supported financially by the Austrian Science Fund
  (FWF, Grant No. I5812-B, “AlloSpace”). This research was supported by the Scientific
  Service Units (SSU) of Institute of Science and Technology Austria (ISTA) through
  resources provided by the Nuclear Magnetic Resonance Facility and the Lab Support
  Facility (LSF). We thank Petra Rovò and Margarita Valhondo Falcón for excellent
  support of the NMR facility.
article_number: '169379'
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Darja
  full_name: Rohden, Darja
  id: 81dc668a-19fa-11f0-bf31-d56534059ef3
  last_name: Rohden
- first_name: Federico
  full_name: Napoli, Federico
  id: d42e08e7-f4fc-11eb-af0a-d71e26138f1b
  last_name: Napoli
  orcid: 0000-0002-9043-136X
- first_name: Anna
  full_name: Kapitonova, Anna
  id: 9fb2a840-89e1-11ee-a8b7-cc5c7ba62471
  last_name: Kapitonova
- first_name: Benjamin
  full_name: Tatman, Benjamin
  id: 71cda2f3-e604-11ee-a1df-da10587eda3f
  last_name: Tatman
- first_name: Roman J.
  full_name: Lichtenecker, Roman J.
  last_name: Lichtenecker
- first_name: Paul
  full_name: Schanda, Paul
  id: 7B541462-FAF6-11E9-A490-E8DFE5697425
  last_name: Schanda
  orcid: 0000-0002-9350-7606
citation:
  ama: Rohden D, Napoli F, Kapitonova A, Tatman B, Lichtenecker RJ, Schanda P. Arginine
    dynamics probed by magic-angle spinning NMR with a specific isotope-labeling scheme.
    <i>Journal of Molecular Biology</i>. 2025;437(23). doi:<a href="https://doi.org/10.1016/j.jmb.2025.169379">10.1016/j.jmb.2025.169379</a>
  apa: Rohden, D., Napoli, F., Kapitonova, A., Tatman, B., Lichtenecker, R. J., &#38;
    Schanda, P. (2025). Arginine dynamics probed by magic-angle spinning NMR with
    a specific isotope-labeling scheme. <i>Journal of Molecular Biology</i>. Elsevier.
    <a href="https://doi.org/10.1016/j.jmb.2025.169379">https://doi.org/10.1016/j.jmb.2025.169379</a>
  chicago: Rohden, Darja, Federico Napoli, Anna Kapitonova, Benjamin Tatman, Roman
    J. Lichtenecker, and Paul Schanda. “Arginine Dynamics Probed by Magic-Angle Spinning
    NMR with a Specific Isotope-Labeling Scheme.” <i>Journal of Molecular Biology</i>.
    Elsevier, 2025. <a href="https://doi.org/10.1016/j.jmb.2025.169379">https://doi.org/10.1016/j.jmb.2025.169379</a>.
  ieee: D. Rohden, F. Napoli, A. Kapitonova, B. Tatman, R. J. Lichtenecker, and P.
    Schanda, “Arginine dynamics probed by magic-angle spinning NMR with a specific
    isotope-labeling scheme,” <i>Journal of Molecular Biology</i>, vol. 437, no. 23.
    Elsevier, 2025.
  ista: Rohden D, Napoli F, Kapitonova A, Tatman B, Lichtenecker RJ, Schanda P. 2025.
    Arginine dynamics probed by magic-angle spinning NMR with a specific isotope-labeling
    scheme. Journal of Molecular Biology. 437(23), 169379.
  mla: Rohden, Darja, et al. “Arginine Dynamics Probed by Magic-Angle Spinning NMR
    with a Specific Isotope-Labeling Scheme.” <i>Journal of Molecular Biology</i>,
    vol. 437, no. 23, 169379, Elsevier, 2025, doi:<a href="https://doi.org/10.1016/j.jmb.2025.169379">10.1016/j.jmb.2025.169379</a>.
  short: D. Rohden, F. Napoli, A. Kapitonova, B. Tatman, R.J. Lichtenecker, P. Schanda,
    Journal of Molecular Biology 437 (2025).
corr_author: '1'
date_created: 2025-08-31T22:01:33Z
date_published: 2025-12-01T00:00:00Z
date_updated: 2025-12-29T14:52:17Z
day: '01'
ddc:
- '540'
department:
- _id: PaSc
doi: 10.1016/j.jmb.2025.169379
external_id:
  isi:
  - '001618289100020'
file:
- access_level: open_access
  checksum: 90d50594d8ea9860ac5da41297992847
  content_type: application/pdf
  creator: dernst
  date_created: 2025-12-29T14:51:40Z
  date_updated: 2025-12-29T14:51:40Z
  file_id: '20876'
  file_name: 2025_JourMolecularBiology_Rohden.pdf
  file_size: 2270555
  relation: main_file
  success: 1
file_date_updated: 2025-12-29T14:51:40Z
has_accepted_license: '1'
intvolume: '       437'
isi: 1
issue: '23'
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
project:
- _id: eb9c82eb-77a9-11ec-83b8-aadd536561cf
  grant_number: I05812
  name: AlloSpace. The emergence and mechanisms of allostery
publication: Journal of Molecular Biology
publication_identifier:
  eissn:
  - 1089-8638
  issn:
  - 0022-2836
publication_status: published
publisher: Elsevier
quality_controlled: '1'
related_material:
  record:
  - id: '19956'
    relation: research_data
    status: public
scopus_import: '1'
status: public
title: Arginine dynamics probed by magic-angle spinning NMR with a specific isotope-labeling
  scheme
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 437
year: '2025'
...
---
OA_place: publisher
OA_type: hybrid
PlanS_conform: '1'
_id: '20538'
abstract:
- lang: eng
  text: In this study, we describe an integrated approach for methyl group assignment
    comprising precursor-based selective methyl group labeling, a novel pulse sequence
    for methyl to backbone coherence transfer and chemical shift predictions using
    UCBShift 2.0. The utility of this novel α-ketoacid isotopologue is shown by the
    adaptation of an HMBC-HMQC pulse sequence that simultaneously connects geminal
    methyl groups of leucine and valine residues to each other and to the protein
    backbone. By additional 13C,2H-labeling of residues other than valine and leucine
    residues of the protein, important chemical shift information about neighboring
    residues (following valine and leucine residues) can be achieved. Thus, different
    valine and leucine residues in a protein can be characterized as a specific chemical
    shift vector. Frequency matching with predicted chemical shifts via UCBShift 2.0
    using experimental data taken from a subset of the BMRB database revealed a correct
    assignment performance of about 90%. With applications to proteins of 60.2 kDa
    and 134 kDa (4 × 33.5 kDa) in size, we demonstrate that the approach provides
    valuable information even for very large proteins.
acknowledged_ssus:
- _id: NMR
- _id: LifeSc
acknowledgement: A.L.P and G.T were funded by the “New Ideas” program by Vienna Doctoral
  School in Chemistry. S.K. was funded by the Austrian Science Fund FWF P35098-B.
  This work was supported financially by the Austrian Science Fund (FWF, grant numbers
  I06223 and I5812-B, “AlloSpace”). This research was supported by the Scientific
  Service Units (SSU) of Institute of Science and Technology Austria (ISTA) through
  resources provided by the Nuclear Magnetic Resonance Facility and the Lab Support
  Facility (LSF). We thank Celina Sailer for assistance with the analysis of the NMR
  spectrum of HsTom70.
article_number: '169465'
article_processing_charge: Yes (in subscription journal)
article_type: original
author:
- first_name: Sonja
  full_name: Knödlstorfer, Sonja
  last_name: Knödlstorfer
- first_name: Giorgia
  full_name: Toscano, Giorgia
  id: 334a5e40-8747-11f0-b671-ba1f5154b4b4
  last_name: Toscano
- first_name: Aleksandra L.
  full_name: Ptaszek, Aleksandra L.
  last_name: Ptaszek
- first_name: Georg
  full_name: Kontaxis, Georg
  last_name: Kontaxis
- first_name: Federico
  full_name: Napoli, Federico
  id: d42e08e7-f4fc-11eb-af0a-d71e26138f1b
  last_name: Napoli
  orcid: 0000-0002-9043-136X
- first_name: Jakob
  full_name: Schneider, Jakob
  id: 64368429-eb97-11eb-a6c2-c980b1f44415
  last_name: Schneider
- first_name: Katharina
  full_name: Maier, Katharina
  last_name: Maier
- first_name: Anna
  full_name: Kapitonova, Anna
  id: 9fb2a840-89e1-11ee-a8b7-cc5c7ba62471
  last_name: Kapitonova
- first_name: Roman J.
  full_name: Lichtenecker, Roman J.
  last_name: Lichtenecker
- first_name: Paul
  full_name: Schanda, Paul
  id: 7B541462-FAF6-11E9-A490-E8DFE5697425
  last_name: Schanda
  orcid: 0000-0002-9350-7606
- first_name: Robert
  full_name: Konrat, Robert
  last_name: Konrat
citation:
  ama: Knödlstorfer S, Toscano G, Ptaszek AL, et al. A novel HMBC-CC-HMQC NMR strategy
    for methyl assignment using triple-13C-labeled α-ketoisovalerate integrated with
    UCBShift 2.0. <i>Journal of Molecular Biology</i>. 2025;437(23). doi:<a href="https://doi.org/10.1016/j.jmb.2025.169465">10.1016/j.jmb.2025.169465</a>
  apa: Knödlstorfer, S., Toscano, G., Ptaszek, A. L., Kontaxis, G., Napoli, F., Schneider,
    J., … Konrat, R. (2025). A novel HMBC-CC-HMQC NMR strategy for methyl assignment
    using triple-13C-labeled α-ketoisovalerate integrated with UCBShift 2.0. <i>Journal
    of Molecular Biology</i>. Elsevier. <a href="https://doi.org/10.1016/j.jmb.2025.169465">https://doi.org/10.1016/j.jmb.2025.169465</a>
  chicago: Knödlstorfer, Sonja, Giorgia Toscano, Aleksandra L. Ptaszek, Georg Kontaxis,
    Federico Napoli, Jakob Schneider, Katharina Maier, et al. “A Novel HMBC-CC-HMQC
    NMR Strategy for Methyl Assignment Using Triple-13C-Labeled α-Ketoisovalerate
    Integrated with UCBShift 2.0.” <i>Journal of Molecular Biology</i>. Elsevier,
    2025. <a href="https://doi.org/10.1016/j.jmb.2025.169465">https://doi.org/10.1016/j.jmb.2025.169465</a>.
  ieee: S. Knödlstorfer <i>et al.</i>, “A novel HMBC-CC-HMQC NMR strategy for methyl
    assignment using triple-13C-labeled α-ketoisovalerate integrated with UCBShift
    2.0,” <i>Journal of Molecular Biology</i>, vol. 437, no. 23. Elsevier, 2025.
  ista: Knödlstorfer S, Toscano G, Ptaszek AL, Kontaxis G, Napoli F, Schneider J,
    Maier K, Kapitonova A, Lichtenecker RJ, Schanda P, Konrat R. 2025. A novel HMBC-CC-HMQC
    NMR strategy for methyl assignment using triple-13C-labeled α-ketoisovalerate
    integrated with UCBShift 2.0. Journal of Molecular Biology. 437(23), 169465.
  mla: Knödlstorfer, Sonja, et al. “A Novel HMBC-CC-HMQC NMR Strategy for Methyl Assignment
    Using Triple-13C-Labeled α-Ketoisovalerate Integrated with UCBShift 2.0.” <i>Journal
    of Molecular Biology</i>, vol. 437, no. 23, 169465, Elsevier, 2025, doi:<a href="https://doi.org/10.1016/j.jmb.2025.169465">10.1016/j.jmb.2025.169465</a>.
  short: S. Knödlstorfer, G. Toscano, A.L. Ptaszek, G. Kontaxis, F. Napoli, J. Schneider,
    K. Maier, A. Kapitonova, R.J. Lichtenecker, P. Schanda, R. Konrat, Journal of
    Molecular Biology 437 (2025).
date_created: 2025-10-26T23:01:35Z
date_published: 2025-12-01T00:00:00Z
date_updated: 2025-12-30T10:29:20Z
day: '01'
ddc:
- '540'
department:
- _id: PaSc
- _id: GradSch
doi: 10.1016/j.jmb.2025.169465
external_id:
  pmid:
  - '41016549'
file:
- access_level: open_access
  checksum: feb92f9c79032c261165f4ca573f444a
  content_type: application/pdf
  creator: dernst
  date_created: 2025-12-30T10:29:08Z
  date_updated: 2025-12-30T10:29:08Z
  file_id: '20915'
  file_name: 2025_JourMolecularBiology_Knoedlstorfer.pdf
  file_size: 3076611
  relation: main_file
  success: 1
file_date_updated: 2025-12-30T10:29:08Z
has_accepted_license: '1'
intvolume: '       437'
issue: '23'
language:
- iso: eng
month: '12'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: bdb9578d-d553-11ed-ba76-ed5d39fce6f0
  grant_number: I06223
  name: Structure and mechanism of the mitochondrial MIM insertase
- _id: eb9c82eb-77a9-11ec-83b8-aadd536561cf
  grant_number: I05812
  name: AlloSpace. The emergence and mechanisms of allostery
publication: Journal of Molecular Biology
publication_identifier:
  eissn:
  - 1089-8638
  issn:
  - 0022-2836
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: A novel HMBC-CC-HMQC NMR strategy for methyl assignment using triple-13C-labeled
  α-ketoisovalerate integrated with UCBShift 2.0
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 437
year: '2025'
...
---
OA_place: publisher
OA_type: hybrid
PlanS_conform: '1'
_id: '19725'
abstract:
- lang: eng
  text: Protein-protein interactions (PPIs) mediate many fundamental cellular processes.
    Control of PPIs through optically or chemically responsive protein domains has
    had a profound impact on basic research and some clinical applications. Most chemogenetic
    methods induce the association, i.e., dimerization or oligomerization, of target
    proteins, whilst the few available dissociation approaches either break large
    oligomeric protein clusters or heteromeric complexes. Here, we have exploited
    the controlled dissociation of a homodimeric oxidoreductase from mycobacteria
    (MSMEG_2027) by its native cofactor, F420, which is not present in mammals, as
    a bioorthogonal monomerization switch. Using X-ray crystallography, we found that
    in the absence of F420 MSMEG_2027 forms a unique domain-swapped dimer that occludes
    the cofactor binding site. Rearrangement of the N-terminal helix upon F420 binding
    results in the dissolution of the dimer. We then showed that MSMEG_2027 can be
    fused to proteins of interest in human cells and applied it as a tool to induce
    and release MAPK/ERK signalling downstream of a chimeric fibroblast growth factor
    receptor 1 (FGFR1) tyrosine kinase. This F420-dependent chemogenetic de-homodimerization
    tool is stoichiometric and based on a single domain and thus represents a novel
    mechanism to investigate protein complexes in situ.
acknowledgement: We thank J. Kaczmarski for advice on isothermal titration calorimetry
  and helpful comments, and Alexandra Tichy, Elliot Gerrard and Rahkesh T Sabapathy
  for assistance with experiments. This study was supported by grants of the Australian
  Research Council (FT200100519 and DP200102093, to H.J.; DE190100806, DP220101901,
  FT230100203, and DP250102939 to T.P.S.D.C; DP200102093, CE200100029 and CE200100012
  to C.J.J.), the National Health and Medical Research Council (APP1187638, to H.J.).
  S.K. was supported by the graduate program MolecularDrugTargets (Austrian Science
  Fund FWF W1232). The Australian Regenerative Medicine Institute is supported by
  grants from the State Government of Victoria and the Australian Government. The
  EMBL Australia Partnership Laboratory (EMBL Australia) is supported by the National
  Collaborative Research Infrastructure Strategy (NCRIS) of the Australian Government.
  T.P.S.D.C. acknowledges the University of Adelaide for a Future Making Fellowship.
  E.R.R.M acknowledges the Grains Research and Development Corporation (9176977) for
  support through a PhD scholarship and operational funding. J.A. and E.R.R.M. were
  supported by Australian Research Training Program scholarship. MicroMon of Monash
  University provided Sanger sequencing services. Imaging was performed in the CellScreen
  SA screening center of Flinders University. C.J.J. thanks the ARC Centre of Excellence
  for Innovations in Peptide and Protein Science and the ARC Centre of Excellence
  in Synthetic Biology. We thank the staff of the MX2 beamline at the Australian Synchrotron,
  part of ANSTO, which made use of the Australian Cancer Research Foundation (ACRF)
  detector.
article_number: '169184'
article_processing_charge: Yes (in subscription journal)
article_type: original
author:
- first_name: James
  full_name: Antoney, James
  last_name: Antoney
- first_name: Stephanie
  full_name: Kainrath, Stephanie
  id: 32CFBA64-F248-11E8-B48F-1D18A9856A87
  last_name: Kainrath
  orcid: 0000-0002-6709-2195
- first_name: Joshua G.
  full_name: Dubowsky, Joshua G.
  last_name: Dubowsky
- first_name: F. Hafna
  full_name: Ahmed, F. Hafna
  last_name: Ahmed
- first_name: Suk Woo
  full_name: Kang, Suk Woo
  last_name: Kang
- first_name: Emily R.R.
  full_name: Mackie, Emily R.R.
  last_name: Mackie
- first_name: Gustavo
  full_name: Bracho Granado, Gustavo
  last_name: Bracho Granado
- first_name: Tatiana P.
  full_name: Soares Da Costa, Tatiana P.
  last_name: Soares Da Costa
- first_name: Colin J.
  full_name: Jackson, Colin J.
  last_name: Jackson
- first_name: Harald L
  full_name: Janovjak, Harald L
  id: 33BA6C30-F248-11E8-B48F-1D18A9856A87
  last_name: Janovjak
  orcid: 0000-0002-8023-9315
citation:
  ama: Antoney J, Kainrath S, Dubowsky JG, et al. A F420-dependent single domain chemogenetic
    tool for protein de-dimerization. <i>Journal of Molecular Biology</i>. 2025;437(17).
    doi:<a href="https://doi.org/10.1016/j.jmb.2025.169184">10.1016/j.jmb.2025.169184</a>
  apa: Antoney, J., Kainrath, S., Dubowsky, J. G., Ahmed, F. H., Kang, S. W., Mackie,
    E. R. R., … Janovjak, H. L. (2025). A F420-dependent single domain chemogenetic
    tool for protein de-dimerization. <i>Journal of Molecular Biology</i>. Elsevier.
    <a href="https://doi.org/10.1016/j.jmb.2025.169184">https://doi.org/10.1016/j.jmb.2025.169184</a>
  chicago: Antoney, James, Stephanie Kainrath, Joshua G. Dubowsky, F. Hafna Ahmed,
    Suk Woo Kang, Emily R.R. Mackie, Gustavo Bracho Granado, Tatiana P. Soares Da
    Costa, Colin J. Jackson, and Harald L Janovjak. “A F420-Dependent Single Domain
    Chemogenetic Tool for Protein de-Dimerization.” <i>Journal of Molecular Biology</i>.
    Elsevier, 2025. <a href="https://doi.org/10.1016/j.jmb.2025.169184">https://doi.org/10.1016/j.jmb.2025.169184</a>.
  ieee: J. Antoney <i>et al.</i>, “A F420-dependent single domain chemogenetic tool
    for protein de-dimerization,” <i>Journal of Molecular Biology</i>, vol. 437, no.
    17. Elsevier, 2025.
  ista: Antoney J, Kainrath S, Dubowsky JG, Ahmed FH, Kang SW, Mackie ERR, Bracho
    Granado G, Soares Da Costa TP, Jackson CJ, Janovjak HL. 2025. A F420-dependent
    single domain chemogenetic tool for protein de-dimerization. Journal of Molecular
    Biology. 437(17), 169184.
  mla: Antoney, James, et al. “A F420-Dependent Single Domain Chemogenetic Tool for
    Protein de-Dimerization.” <i>Journal of Molecular Biology</i>, vol. 437, no. 17,
    169184, Elsevier, 2025, doi:<a href="https://doi.org/10.1016/j.jmb.2025.169184">10.1016/j.jmb.2025.169184</a>.
  short: J. Antoney, S. Kainrath, J.G. Dubowsky, F.H. Ahmed, S.W. Kang, E.R.R. Mackie,
    G. Bracho Granado, T.P. Soares Da Costa, C.J. Jackson, H.L. Janovjak, Journal
    of Molecular Biology 437 (2025).
date_created: 2025-05-25T22:16:39Z
date_published: 2025-09-01T00:00:00Z
date_updated: 2025-12-30T08:18:25Z
day: '01'
ddc:
- '570'
department:
- _id: CaGu
doi: 10.1016/j.jmb.2025.169184
external_id:
  isi:
  - '001494762800001'
  pmid:
  - '40324743'
file:
- access_level: open_access
  checksum: fb6e84ba7dc92faee97647fd2bc8cca8
  content_type: application/pdf
  creator: dernst
  date_created: 2025-12-30T08:18:07Z
  date_updated: 2025-12-30T08:18:07Z
  file_id: '20892'
  file_name: 2025_JourMolecularBiology_Antoney.pdf
  file_size: 1682721
  relation: main_file
  success: 1
file_date_updated: 2025-12-30T08:18:07Z
has_accepted_license: '1'
intvolume: '       437'
isi: 1
issue: '17'
language:
- iso: eng
month: '09'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 255A6082-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: W1232-B24
  name: Molecular Drug Targets
publication: Journal of Molecular Biology
publication_identifier:
  eissn:
  - 1089-8638
  issn:
  - 0022-2836
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: A F420-dependent single domain chemogenetic tool for protein de-dimerization
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 437
year: '2025'
...
---
_id: '6564'
abstract:
- lang: eng
  text: Optogenetics enables the spatio-temporally precise control of cell and animal
    behavior. Many optogenetic tools are driven by light-controlled protein–protein
    interactions (PPIs) that are repurposed from natural light-sensitive domains (LSDs).
    Applying light-controlled PPIs to new target proteins is challenging because it
    is difficult to predict which of the many available LSDs, if any, will yield robust
    light regulation. As a consequence, fusion protein libraries need to be prepared
    and tested, but methods and platforms to facilitate this process are currently
    not available. Here, we developed a genetic engineering strategy and vector library
    for the rapid generation of light-controlled PPIs. The strategy permits fusing
    a target protein to multiple LSDs efficiently and in two orientations. The public
    and expandable library contains 29 vectors with blue, green or red light-responsive
    LSDs, many of which have been previously applied ex vivo and in vivo. We demonstrate
    the versatility of the approach and the necessity for sampling LSDs by generating
    light-activated caspase-9 (casp9) enzymes. Collectively, this work provides a
    new resource for optical regulation of a broad range of target proteins in cell
    and developmental biology.
article_processing_charge: No
article_type: original
author:
- first_name: Alexandra-Madelaine
  full_name: Tichy, Alexandra-Madelaine
  id: 29D8BB2C-F248-11E8-B48F-1D18A9856A87
  last_name: Tichy
- first_name: Elliot J.
  full_name: Gerrard, Elliot J.
  last_name: Gerrard
- first_name: Julien M.D.
  full_name: Legrand, Julien M.D.
  last_name: Legrand
- first_name: Robin M.
  full_name: Hobbs, Robin M.
  last_name: Hobbs
- first_name: Harald L
  full_name: Janovjak, Harald L
  id: 33BA6C30-F248-11E8-B48F-1D18A9856A87
  last_name: Janovjak
  orcid: 0000-0002-8023-9315
citation:
  ama: Tichy A-M, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak HL. Engineering strategy
    and vector library for the rapid generation of modular light-controlled protein–protein
    interactions. <i>Journal of Molecular Biology</i>. 2019;431(17):3046-3055. doi:<a
    href="https://doi.org/10.1016/j.jmb.2019.05.033">10.1016/j.jmb.2019.05.033</a>
  apa: Tichy, A.-M., Gerrard, E. J., Legrand, J. M. D., Hobbs, R. M., &#38; Janovjak,
    H. L. (2019). Engineering strategy and vector library for the rapid generation
    of modular light-controlled protein–protein interactions. <i>Journal of Molecular
    Biology</i>. Elsevier. <a href="https://doi.org/10.1016/j.jmb.2019.05.033">https://doi.org/10.1016/j.jmb.2019.05.033</a>
  chicago: Tichy, Alexandra-Madelaine, Elliot J. Gerrard, Julien M.D. Legrand, Robin
    M. Hobbs, and Harald L Janovjak. “Engineering Strategy and Vector Library for
    the Rapid Generation of Modular Light-Controlled Protein–Protein Interactions.”
    <i>Journal of Molecular Biology</i>. Elsevier, 2019. <a href="https://doi.org/10.1016/j.jmb.2019.05.033">https://doi.org/10.1016/j.jmb.2019.05.033</a>.
  ieee: A.-M. Tichy, E. J. Gerrard, J. M. D. Legrand, R. M. Hobbs, and H. L. Janovjak,
    “Engineering strategy and vector library for the rapid generation of modular light-controlled
    protein–protein interactions,” <i>Journal of Molecular Biology</i>, vol. 431,
    no. 17. Elsevier, pp. 3046–3055, 2019.
  ista: Tichy A-M, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak HL. 2019. Engineering
    strategy and vector library for the rapid generation of modular light-controlled
    protein–protein interactions. Journal of Molecular Biology. 431(17), 3046–3055.
  mla: Tichy, Alexandra-Madelaine, et al. “Engineering Strategy and Vector Library
    for the Rapid Generation of Modular Light-Controlled Protein–Protein Interactions.”
    <i>Journal of Molecular Biology</i>, vol. 431, no. 17, Elsevier, 2019, pp. 3046–55,
    doi:<a href="https://doi.org/10.1016/j.jmb.2019.05.033">10.1016/j.jmb.2019.05.033</a>.
  short: A.-M. Tichy, E.J. Gerrard, J.M.D. Legrand, R.M. Hobbs, H.L. Janovjak, Journal
    of Molecular Biology 431 (2019) 3046–3055.
date_created: 2019-06-16T21:59:14Z
date_published: 2019-08-09T00:00:00Z
date_updated: 2025-07-10T11:53:33Z
day: '09'
department:
- _id: HaJa
doi: 10.1016/j.jmb.2019.05.033
external_id:
  isi:
  - '000482872100002'
intvolume: '       431'
isi: 1
issue: '17'
language:
- iso: eng
main_file_link:
- open_access: '1'
  url: http://www.biorxiv.org/content/10.1101/583369v1
month: '08'
oa: 1
oa_version: Preprint
page: 3046-3055
publication: Journal of Molecular Biology
publication_identifier:
  eissn:
  - 1089-8638
  issn:
  - 0022-2836
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Engineering strategy and vector library for the rapid generation of modular
  light-controlled protein–protein interactions
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 431
year: '2019'
...