---
OA_place: publisher
OA_type: hybrid
PlanS_conform: '1'
_id: '21929'
abstract:
- lang: eng
text: 'The import of proteins into mitochondria poses fundamental mechanistic challenges:
aggregation-prone precursor proteins must be maintained in aqueous compartments
and threaded through narrow pores without becoming stuck or mislocalized. Recent
evidence from mitochondrial protein import studies and other chaperone systems
underscores the critical role of dynamics in balancing sufficiently tight binding,
promiscuity, specificity, and release. Dynamic binding of client precursor proteins
to import machinery components arises naturally from the avidity of their interactions.
Conformational entropy enhances their stability, while the multivalent nature
of these interactions ensures that client transfer to downstream insertases occurs
without a substantial energy barrier. Here, we discuss this emerging paradigm
of dynamic protein handling, using examples where dynamic structures have been
resolved and highlight outstanding questions.'
acknowledgement: We gratefully acknowledge research funding by the Austrian Science
Fund (FWF), projects 10.55776/PAT1647625 and 10.55776/I6223. We thank Prof. Long
Li (Peking University) for providing structural models and EM density for the TOM
and TIM23 complexes, used to generate part of Figure 3. Open Access funding provided
by Institute of Science and Technology Austria.
article_number: e70630
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Jakob
full_name: Schneider, Jakob
id: 64368429-eb97-11eb-a6c2-c980b1f44415
last_name: Schneider
- first_name: Undina
full_name: Guillerm, Undina
id: bb74f472-ae54-11eb-9835-bc9c22fb1183
last_name: Guillerm
- first_name: Caroline
full_name: Simoes Pereira, Caroline
id: 87266c4a-96d2-11ef-be2c-fe5633233ec3
last_name: Simoes Pereira
- first_name: Paul
full_name: Schanda, Paul
id: 7B541462-FAF6-11E9-A490-E8DFE5697425
last_name: Schanda
orcid: 0000-0002-9350-7606
citation:
ama: Schneider J, Guillerm U, Simoes Pereira C, Schanda P. Dynamic disorder is crucial
for mitochondrial protein import. Protein Science. 2026;35(6). doi:10.1002/pro.70630
apa: Schneider, J., Guillerm, U., Simoes Pereira, C., & Schanda, P. (2026).
Dynamic disorder is crucial for mitochondrial protein import. Protein Science.
Wiley. https://doi.org/10.1002/pro.70630
chicago: Schneider, Jakob, Undina Guillerm, Caroline Simoes Pereira, and Paul Schanda.
“Dynamic Disorder Is Crucial for Mitochondrial Protein Import.” Protein Science.
Wiley, 2026. https://doi.org/10.1002/pro.70630.
ieee: J. Schneider, U. Guillerm, C. Simoes Pereira, and P. Schanda, “Dynamic disorder
is crucial for mitochondrial protein import,” Protein Science, vol. 35,
no. 6. Wiley, 2026.
ista: Schneider J, Guillerm U, Simoes Pereira C, Schanda P. 2026. Dynamic disorder
is crucial for mitochondrial protein import. Protein Science. 35(6), e70630.
mla: Schneider, Jakob, et al. “Dynamic Disorder Is Crucial for Mitochondrial Protein
Import.” Protein Science, vol. 35, no. 6, e70630, Wiley, 2026, doi:10.1002/pro.70630.
short: J. Schneider, U. Guillerm, C. Simoes Pereira, P. Schanda, Protein Science
35 (2026).
corr_author: '1'
date_created: 2026-05-31T22:02:12Z
date_published: 2026-06-01T00:00:00Z
date_updated: 2026-06-02T07:26:34Z
day: '01'
ddc:
- '572'
department:
- _id: GradSch
- _id: PaSc
doi: 10.1002/pro.70630
external_id:
pmid:
- '42159315'
file:
- access_level: open_access
checksum: e0163459a7238fdcc3fc5e17bedcce9a
content_type: application/pdf
creator: dernst
date_created: 2026-06-02T07:23:12Z
date_updated: 2026-06-02T07:23:12Z
file_id: '21937'
file_name: 2026_ProteinScience_Schneider.pdf
file_size: 3897305
relation: main_file
success: 1
file_date_updated: 2026-06-02T07:23:12Z
fulldoi: https://doi.org/10.1002/pro.70630
has_accepted_license: '1'
intvolume: ' 35'
issue: '6'
language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '06'
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
publication: Protein Science
publication_identifier:
eissn:
- 1469-896X
issn:
- 0961-8368
publication_status: published
publisher: Wiley
quality_controlled: '1'
scopus_import: '1'
status: public
title: Dynamic disorder is crucial for mitochondrial protein import
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: 35
year: '2026'
...
---
DOAJ_listed: '1'
OA_place: publisher
OA_type: gold
PlanS_conform: '1'
_id: '21777'
abstract:
- lang: eng
text: The advantageous characteristics attributed to the 19F nucleus have made it
a popular target for nuclear magnetic resonance (NMR) once again in recent years.
Aside from solution NMR, an increasing number of studies have been conducted applying
solid-state magic-angle spinning (MAS) NMR to fluorine-labelled samples. Here,
the high chemical shift anisotropy and strong dipolar couplings can be utilised
to get structural insights into proteins and measure long distances. Despite increasing
popularity and promising benefits, the sensitivity of biomolecular 19F MAS NMR
often suffers from slow longitudinal T1 relaxation and therefore long recycle
delays. In this work, we expand paramagnetic doping, an approach commonly used
to reduce proton T1 relaxation times, to 19F-labelled biological samples. We study
the effect of Gd(DTPA) and Gd(DTPA-BMA) on 19F T1 and T2, and 13C T1 and T2 relaxation
in a [5-19F13C]-tryptophan-labelled protein via 19F-detected MAS NMR experiments.
The observed paramagnetic relaxation enhancement substantially reduces measurement
times of 19F MAS NMR experiments without compromising resolution. Additionally,
we report the chemical shift assignments of all four fluorotryptophan signals
in the 12×39 kDa-large protein TET2 using a mutagenesis approach.
acknowledged_ssus:
- _id: NMR
- _id: LifeSc
acknowledgement: We thank Ben P. Tatman for insightful discussions. This research
was supported by the Scientific Service Units (SSUs) of ISTA through resources provided
by the Nuclear Magnetic Resonance Facility and the Lab Support Facility. We thank
Prof. Tobias Madl (Medical University Graz) for a sample of Omniscan. Lea M. Becker
is the recipient of a DOC fellowship of the Austrian Academy of Sciences at the
Institute of Science and Technology Austria (grant no. PR10660EAW01).
article_processing_charge: Yes
article_type: original
author:
- first_name: Lea Marie
full_name: Becker, Lea Marie
id: 36336939-eb97-11eb-a6c2-c83f1214ca79
last_name: Becker
orcid: 0000-0002-6401-5151
- first_name: Giorgia
full_name: Toscano, Giorgia
id: 334a5e40-8747-11f0-b671-ba1f5154b4b4
last_name: Toscano
- first_name: Anna
full_name: Kapitonova, Anna
id: 9fb2a840-89e1-11ee-a8b7-cc5c7ba62471
last_name: Kapitonova
- first_name: Rajkumar
full_name: Singh, Rajkumar
id: a3089acd-6806-11ee-bacc-f0c7d500ad20
last_name: Singh
- first_name: Undina
full_name: Guillerm, Undina
id: bb74f472-ae54-11eb-9835-bc9c22fb1183
last_name: Guillerm
- 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: Becker LM, Toscano G, Kapitonova A, et al. Accelerated 19F biomolecular magic-angle
spinning NMR with paramagnetic dopants. Magnetic Resonance. 2026;7(1):29-37.
doi:10.5194/mr-7-29-2026
apa: Becker, L. M., Toscano, G., Kapitonova, A., Singh, R., Guillerm, U., Lichtenecker,
R. J., & Schanda, P. (2026). Accelerated 19F biomolecular magic-angle spinning
NMR with paramagnetic dopants. Magnetic Resonance. Copernicus Publications.
https://doi.org/10.5194/mr-7-29-2026
chicago: Becker, Lea Marie, Giorgia Toscano, Anna Kapitonova, Rajkumar Singh, Undina
Guillerm, Roman J. Lichtenecker, and Paul Schanda. “Accelerated 19F Biomolecular
Magic-Angle Spinning NMR with Paramagnetic Dopants.” Magnetic Resonance.
Copernicus Publications, 2026. https://doi.org/10.5194/mr-7-29-2026.
ieee: L. M. Becker et al., “Accelerated 19F biomolecular magic-angle spinning
NMR with paramagnetic dopants,” Magnetic Resonance, vol. 7, no. 1. Copernicus
Publications, pp. 29–37, 2026.
ista: Becker LM, Toscano G, Kapitonova A, Singh R, Guillerm U, Lichtenecker RJ,
Schanda P. 2026. Accelerated 19F biomolecular magic-angle spinning NMR with paramagnetic
dopants. Magnetic Resonance. 7(1), 29–37.
mla: Becker, Lea Marie, et al. “Accelerated 19F Biomolecular Magic-Angle Spinning
NMR with Paramagnetic Dopants.” Magnetic Resonance, vol. 7, no. 1, Copernicus
Publications, 2026, pp. 29–37, doi:10.5194/mr-7-29-2026.
short: L.M. Becker, G. Toscano, A. Kapitonova, R. Singh, U. Guillerm, R.J. Lichtenecker,
P. Schanda, Magnetic Resonance 7 (2026) 29–37.
corr_author: '1'
date_created: 2026-05-03T22:01:36Z
date_published: 2026-04-16T00:00:00Z
date_updated: 2026-07-20T09:49:12Z
day: '16'
ddc:
- '540'
department:
- _id: PaSc
- _id: GradSch
doi: 10.5194/mr-7-29-2026
external_id:
pmid:
- '42057802'
fulldoi: https://doi.org/10.5194/mr-7-29-2026
has_accepted_license: '1'
intvolume: ' 7'
issue: '1'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://doi.org/10.5194/mr-7-29-2026
month: '04'
oa: 1
oa_version: Published Version
page: 29-37
pmid: 1
project:
- _id: B67AFEDC-15C9-11EA-A837-991A96BB2854
name: IST Austria Open Access Fund
- _id: 7be609c4-9f16-11ee-852c-85015ce2b9b0
grant_number: '26777'
name: Exploring protein dynamics by solid-state MAS NMR through specific labeling
approaches
publication: Magnetic Resonance
publication_identifier:
eissn:
- 2699-0016
publication_status: published
publisher: Copernicus Publications
quality_controlled: '1'
related_material:
record:
- id: '22334'
relation: dissertation_contains
status: public
scopus_import: '1'
status: public
title: Accelerated 19F biomolecular magic-angle spinning NMR with paramagnetic dopants
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: 7
year: '2026'
...
---
OA_type: closed access
_id: '18167'
abstract:
- lang: eng
text: 'Holdase chaperones are essential in the mitochondrial membrane-protein biogenesis
as they stabilize preproteins and keep them in an import-competent state as they
travel through the aqueous cytosol and intermembrane space. The small TIM chaperones
of the mitochondrial intermembrane space function within a fine balance of client
promiscuity and high affinity binding, while being also able to release their
client proteins without significant energy barrier to the downstream insertases/translocases.
The tendency of the preproteins to aggregate and the dynamic nature of the preprotein—chaperone
complexes makes the preparation of these complexes challenging. Here we present
two optimized methods for complex formation of highly hydrophobic precursor proteins
and chaperones: a pull-down approach and an in-vitro translation strategy. In
the former, attaching the client protein to an affinity resin keeps the individual
client protein copies apart from each other and decreases the client self-aggregation
probability, thereby favouring complex formation. In the latter approach, a purified
chaperone, added to the cell-free protein synthesis, captures the nascent precursor
protein. The choice of method will depend on the desired client-chaperone complex
amount, or the need for specific labeling scheme.'
article_processing_charge: No
author:
- first_name: Undina
full_name: Guillerm, Undina
id: bb74f472-ae54-11eb-9835-bc9c22fb1183
last_name: Guillerm
- first_name: Iva
full_name: Sučec, Iva
last_name: Sučec
- first_name: Paul
full_name: Schanda, Paul
id: 7B541462-FAF6-11E9-A490-E8DFE5697425
last_name: Schanda
orcid: 0000-0002-9350-7606
citation:
ama: 'Guillerm U, Sučec I, Schanda P. Generation of TIM chaperone substrate complexes.
In: Methods in Enzymology. Vol 707. Elsevier; 2024:391-422. doi:10.1016/bs.mie.2024.07.051'
apa: Guillerm, U., Sučec, I., & Schanda, P. (2024). Generation of TIM chaperone
substrate complexes. In Methods in Enzymology (Vol. 707, pp. 391–422).
Elsevier. https://doi.org/10.1016/bs.mie.2024.07.051
chicago: Guillerm, Undina, Iva Sučec, and Paul Schanda. “Generation of TIM Chaperone
Substrate Complexes.” In Methods in Enzymology, 707:391–422. Elsevier,
2024. https://doi.org/10.1016/bs.mie.2024.07.051.
ieee: U. Guillerm, I. Sučec, and P. Schanda, “Generation of TIM chaperone substrate
complexes,” in Methods in Enzymology, vol. 707, Elsevier, 2024, pp. 391–422.
ista: 'Guillerm U, Sučec I, Schanda P. 2024.Generation of TIM chaperone substrate
complexes. In: Methods in Enzymology. vol. 707, 391–422.'
mla: Guillerm, Undina, et al. “Generation of TIM Chaperone Substrate Complexes.”
Methods in Enzymology, vol. 707, Elsevier, 2024, pp. 391–422, doi:10.1016/bs.mie.2024.07.051.
short: U. Guillerm, I. Sučec, P. Schanda, in:, Methods in Enzymology, Elsevier,
2024, pp. 391–422.
corr_author: '1'
date_created: 2024-10-01T10:58:27Z
date_published: 2024-09-13T00:00:00Z
date_updated: 2025-10-22T06:40:54Z
day: '13'
department:
- _id: PaSc
doi: 10.1016/bs.mie.2024.07.051
external_id:
pmid:
- '39488384'
fulldoi: https://doi.org/10.1016/bs.mie.2024.07.051
intvolume: ' 707'
language:
- iso: eng
month: '09'
oa_version: None
page: 391-422
pmid: 1
publication: Methods in Enzymology
publication_identifier:
issn:
- 0076-6879
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Generation of TIM chaperone substrate complexes
type: book_chapter
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 707
year: '2024'
...
---
_id: '13096'
abstract:
- lang: eng
text: Eukaryotic cells can undergo different forms of programmed cell death, many
of which culminate in plasma membrane rupture as the defining terminal event1,2,3,4,5,6,7.
Plasma membrane rupture was long thought to be driven by osmotic pressure, but
it has recently been shown to be in many cases an active process, mediated by
the protein ninjurin-18 (NINJ1). Here we resolve the structure of NINJ1 and the
mechanism by which it ruptures membranes. Super-resolution microscopy reveals
that NINJ1 clusters into structurally diverse assemblies in the membranes of dying
cells, in particular large, filamentous assemblies with branched morphology. A
cryo-electron microscopy structure of NINJ1 filaments shows a tightly packed fence-like
array of transmembrane α-helices. Filament directionality and stability is defined
by two amphipathic α-helices that interlink adjacent filament subunits. The NINJ1
filament features a hydrophilic side and a hydrophobic side, and molecular dynamics
simulations show that it can stably cap membrane edges. The function of the resulting
supramolecular arrangement was validated by site-directed mutagenesis. Our data
thus suggest that, during lytic cell death, the extracellular α-helices of NINJ1
insert into the plasma membrane to polymerize NINJ1 monomers into amphipathic
filaments that rupture the plasma membrane. The membrane protein NINJ1 is therefore
an interactive component of the eukaryotic cell membrane that functions as an
in-built breaking point in response to activation of cell death.
acknowledged_ssus:
- _id: NMR
- _id: LifeSc
acknowledgement: "This work was supported by the Deutsche Forschungsgemeinschaft under
Germany’s Excellence Strategy EXC 2075–390740016 and the Stuttgart Center for Simulation
Science (SC SimTech) to K.P., by ERC-CoG 770988 (InflamCellDeath) and SNF Project
funding (310030B_198005, 310030B_192523) to P.B., by the Swiss Nanoscience Institute
and the Swiss National Science Foundation via the NCCR AntiResist (180541) to S.H.
and the NCCR Molecular Systems Engineering (51NF40-205608) to D.J.M., by the Helmholtz
Young Investigator Program of the Helmholtz Association to C.S., by the SNF Professorship
funding (PP00P3_198903) to C.P., EMBO postdoctoral fellowship ALTF 27-2022 to E.H.
and by the Scientific Service Units of IST Austria through resources provided by
the NMR and Life Science Facilities to P.S. Molecular dynamics simulations were
performed on the HoreKa supercomputer funded by the Ministry of Science, Research
and the Arts Baden-Württemberg and by the Federal Ministry of Education and Research.
The authors thank the BioEM Lab of the Biozentrum, University of Basel for support;
V. Mack, K. Shkarina and J. Fricke for technical support; D. Ricklin and S. Vogt
for peptide synthesis; P. Pelczar for support with animals; S.-J. Marrink and P.
Telles de Souza for supply with Martini3 parameters and scripts; and P. Radler und
M. Loose for help with QCM. Fig. 4g and Extended Data Fig. 1a were in part created
with BioRender.com.\r\nOpen access funding provided by University of Basel."
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Morris
full_name: Degen, Morris
last_name: Degen
- first_name: José Carlos
full_name: Santos, José Carlos
last_name: Santos
- first_name: Kristyna
full_name: Pluhackova, Kristyna
last_name: Pluhackova
- first_name: Gonzalo
full_name: Cebrero, Gonzalo
last_name: Cebrero
- first_name: Saray
full_name: Ramos, Saray
last_name: Ramos
- first_name: Gytis
full_name: Jankevicius, Gytis
last_name: Jankevicius
- first_name: Ella
full_name: Hartenian, Ella
last_name: Hartenian
- first_name: Undina
full_name: Guillerm, Undina
id: bb74f472-ae54-11eb-9835-bc9c22fb1183
last_name: Guillerm
- first_name: Stefania A.
full_name: Mari, Stefania A.
last_name: Mari
- first_name: Bastian
full_name: Kohl, Bastian
last_name: Kohl
- first_name: Daniel J.
full_name: Müller, Daniel J.
last_name: Müller
- first_name: Paul
full_name: Schanda, Paul
id: 7B541462-FAF6-11E9-A490-E8DFE5697425
last_name: Schanda
orcid: 0000-0002-9350-7606
- first_name: Timm
full_name: Maier, Timm
last_name: Maier
- first_name: Camilo
full_name: Perez, Camilo
last_name: Perez
- first_name: Christian
full_name: Sieben, Christian
last_name: Sieben
- first_name: Petr
full_name: Broz, Petr
last_name: Broz
- first_name: Sebastian
full_name: Hiller, Sebastian
last_name: Hiller
citation:
ama: Degen M, Santos JC, Pluhackova K, et al. Structural basis of NINJ1-mediated
plasma membrane rupture in cell death. Nature. 2023;618:1065-1071. doi:10.1038/s41586-023-05991-z
apa: Degen, M., Santos, J. C., Pluhackova, K., Cebrero, G., Ramos, S., Jankevicius,
G., … Hiller, S. (2023). Structural basis of NINJ1-mediated plasma membrane rupture
in cell death. Nature. Springer Nature. https://doi.org/10.1038/s41586-023-05991-z
chicago: Degen, Morris, José Carlos Santos, Kristyna Pluhackova, Gonzalo Cebrero,
Saray Ramos, Gytis Jankevicius, Ella Hartenian, et al. “Structural Basis of NINJ1-Mediated
Plasma Membrane Rupture in Cell Death.” Nature. Springer Nature, 2023.
https://doi.org/10.1038/s41586-023-05991-z.
ieee: M. Degen et al., “Structural basis of NINJ1-mediated plasma membrane
rupture in cell death,” Nature, vol. 618. Springer Nature, pp. 1065–1071,
2023.
ista: Degen M, Santos JC, Pluhackova K, Cebrero G, Ramos S, Jankevicius G, Hartenian
E, Guillerm U, Mari SA, Kohl B, Müller DJ, Schanda P, Maier T, Perez C, Sieben
C, Broz P, Hiller S. 2023. Structural basis of NINJ1-mediated plasma membrane
rupture in cell death. Nature. 618, 1065–1071.
mla: Degen, Morris, et al. “Structural Basis of NINJ1-Mediated Plasma Membrane Rupture
in Cell Death.” Nature, vol. 618, Springer Nature, 2023, pp. 1065–71, doi:10.1038/s41586-023-05991-z.
short: M. Degen, J.C. Santos, K. Pluhackova, G. Cebrero, S. Ramos, G. Jankevicius,
E. Hartenian, U. Guillerm, S.A. Mari, B. Kohl, D.J. Müller, P. Schanda, T. Maier,
C. Perez, C. Sieben, P. Broz, S. Hiller, Nature 618 (2023) 1065–1071.
date_created: 2023-05-28T22:01:04Z
date_published: 2023-06-29T00:00:00Z
date_updated: 2025-04-23T08:57:12Z
day: '29'
ddc:
- '570'
department:
- _id: PaSc
doi: 10.1038/s41586-023-05991-z
external_id:
isi:
- '000991386800011'
pmid:
- '37198476'
file:
- access_level: open_access
checksum: 0fab69252453bff1de7f0e2eceb76d34
content_type: application/pdf
creator: dernst
date_created: 2023-11-14T11:48:18Z
date_updated: 2023-11-14T11:48:18Z
file_id: '14533'
file_name: 2023_Nature_Degen.pdf
file_size: 12292188
relation: main_file
success: 1
file_date_updated: 2023-11-14T11:48:18Z
fulldoi: https://doi.org/10.1038/s41586-023-05991-z
has_accepted_license: '1'
intvolume: ' 618'
isi: 1
language:
- iso: eng
month: '06'
oa: 1
oa_version: Published Version
page: 1065-1071
pmid: 1
publication: Nature
publication_identifier:
eissn:
- 1476-4687
issn:
- 0028-0836
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
status: public
title: Structural basis of NINJ1-mediated plasma membrane rupture in cell death
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: 618
year: '2023'
...