---
OA_place: publisher
OA_type: hybrid
PlanS_conform: '1'
_id: '20404'
abstract:
- lang: eng
text: Collagens are fundamental components of extracellular matrices, requiring
precise intracellular post-translational modifications for proper function. Among
the modifications, prolyl 4-hydroxylation is critical to stabilise the collagen
triple helix. In humans, this reaction is mediated by collagen prolyl 4-hydroxylases
(P4Hs). While humans possess three genes encoding these enzymes (P4H⍺s), Drosophila
melanogaster harbour at least 26 candidates for collagen P4H⍺s despite its simple
genome, and it is poorly understood which of them are actually working on collagen
in the fly. In this study, we addressed this question by carrying out thorough
bioinformatic and biochemical analyses. We demonstrate that among the 26 potential
collagen P4H⍺s, PH4⍺EFB shares the highest homology with vertebrate collagen P4H⍺s.
Furthermore, while collagen P4Hs and their substrates must exist in the same cells,
our transcriptomic analyses at the tissue and single cell levels showed a global
co-expression of PH4⍺EFB but not the other P4H⍺-related genes with the collagen
IV genes. Moreover, expression of PH4⍺EFB during embryogenesis was found to precede
that of collagen IV, presumably enabling efficient collagen modification by PH4⍺EFB.
Finally, biochemical assays confirm that PH4⍺EFB binds collagen, supporting its
direct role in collagen IV modification. Collectively, we identify PH4⍺EFB as
the primary and potentially constitutive prolyl 4-hydroxylase responsible for
collagen IV biosynthesis in Drosophila. Our findings highlight the remarkably
simple nature of Drosophila collagen IV biosynthesis, which may serve as a blueprint
for defining the minimal requirements for collagen engineering.
acknowledgement: "This project was supported by the All May See Foundation 7031,182
to YI, the Louisiana Board of Regents Support Fund: Research Competitiveness Subprogram
to MAT, Austrian science fund (FWF) as part of the SFB Meiosis consortium FWF SFB
F88-10 to Beatriz Vicoso (supported ME), American Heart Association 16POST2726018
and American Cancer Society 132,123-PF-18–025–01-CSM postdoctoral fellowships to
ALZ, National Institutes of Health R01 GM136961 and R35 GM148485 to SH-B, and the
Academy of Medical Sciences/the Wellcome Trust/ the Government Department of Business,
Energy and Industrial Strategy/the British Heart Foundation/Diabetes UK Springboard
Award SBF008\\1115 to YM. \r\nComputational analyses of single-nucleus transcriptome
data were performed on the high performance computer (HPC) at Bournemouth University,
the HPC at Institute of Science and Technology Austria, and the high-performance
computational resources provided by the Louisiana Optical Network Infrastructure
(http://www.loni.org). The authors are grateful to the researchers who published
the transcriptome datasets [48,49,52,55] that became the essential bases for this
study, to FlyBase for curating the datasets in an easily accessible format, and
the Drosophila Genomics Resource Center (DGRC), supported by NIH grant 2P40OD010949,
for providing the D17 cell line used in this research. The authors thank Kristian
Koski (University of Oulu, Finland) for crucial advice on the domain structure of
collagen P4H⍺s, and Ryusuke Niwa and Ryo Hoshino (University of Tsukuba, Japan)
for helpful discussions on SP."
article_processing_charge: Yes (in subscription journal)
article_type: original
author:
- first_name: Yoshihiro
full_name: Ishikawa, Yoshihiro
last_name: Ishikawa
- first_name: Melissa A
full_name: Toups, Melissa A
id: 4E099E4E-F248-11E8-B48F-1D18A9856A87
last_name: Toups
orcid: 0000-0002-9752-7380
- first_name: Marwan N
full_name: Elkrewi, Marwan N
id: 0B46FACA-A8E1-11E9-9BD3-79D1E5697425
last_name: Elkrewi
orcid: 0000-0002-5328-7231
- first_name: Allison L.
full_name: Zajac, Allison L.
last_name: Zajac
- first_name: Sally
full_name: Horne-Badovinac, Sally
last_name: Horne-Badovinac
- first_name: Yutaka
full_name: Matsubayashi, Yutaka
last_name: Matsubayashi
citation:
ama: Ishikawa Y, Toups MA, Elkrewi MN, Zajac AL, Horne-Badovinac S, Matsubayashi
Y. Evidence for the major role of PH4⍺EFB in the prolyl 4-hydroxylation of Drosophila
collagen IV. Matrix Biology. 2025;141(11):101-113. doi:10.1016/j.matbio.2025.09.002
apa: Ishikawa, Y., Toups, M. A., Elkrewi, M. N., Zajac, A. L., Horne-Badovinac,
S., & Matsubayashi, Y. (2025). Evidence for the major role of PH4⍺EFB in the
prolyl 4-hydroxylation of Drosophila collagen IV. Matrix Biology. Springer
Nature. https://doi.org/10.1016/j.matbio.2025.09.002
chicago: Ishikawa, Yoshihiro, Melissa A Toups, Marwan N Elkrewi, Allison L. Zajac,
Sally Horne-Badovinac, and Yutaka Matsubayashi. “Evidence for the Major Role of
PH4⍺EFB in the Prolyl 4-Hydroxylation of Drosophila Collagen IV.” Matrix Biology.
Springer Nature, 2025. https://doi.org/10.1016/j.matbio.2025.09.002.
ieee: Y. Ishikawa, M. A. Toups, M. N. Elkrewi, A. L. Zajac, S. Horne-Badovinac,
and Y. Matsubayashi, “Evidence for the major role of PH4⍺EFB in the prolyl 4-hydroxylation
of Drosophila collagen IV,” Matrix Biology, vol. 141, no. 11. Springer
Nature, pp. 101–113, 2025.
ista: Ishikawa Y, Toups MA, Elkrewi MN, Zajac AL, Horne-Badovinac S, Matsubayashi
Y. 2025. Evidence for the major role of PH4⍺EFB in the prolyl 4-hydroxylation
of Drosophila collagen IV. Matrix Biology. 141(11), 101–113.
mla: Ishikawa, Yoshihiro, et al. “Evidence for the Major Role of PH4⍺EFB in the
Prolyl 4-Hydroxylation of Drosophila Collagen IV.” Matrix Biology, vol.
141, no. 11, Springer Nature, 2025, pp. 101–13, doi:10.1016/j.matbio.2025.09.002.
short: Y. Ishikawa, M.A. Toups, M.N. Elkrewi, A.L. Zajac, S. Horne-Badovinac, Y.
Matsubayashi, Matrix Biology 141 (2025) 101–113.
date_created: 2025-09-28T22:01:26Z
date_published: 2025-11-01T00:00:00Z
date_updated: 2026-01-05T13:09:08Z
day: '01'
ddc:
- '570'
department:
- _id: BeVi
doi: 10.1016/j.matbio.2025.09.002
external_id:
isi:
- '001583892100002'
pmid:
- '40946811'
file:
- access_level: open_access
checksum: 764257db41865d19daec1935788f72d7
content_type: application/pdf
creator: dernst
date_created: 2026-01-05T13:09:01Z
date_updated: 2026-01-05T13:09:01Z
file_id: '20948'
file_name: 2025_MatrixBiology_Ishikawa.pdf
file_size: 5844254
relation: main_file
success: 1
file_date_updated: 2026-01-05T13:09:01Z
has_accepted_license: '1'
intvolume: ' 141'
isi: 1
issue: '11'
language:
- iso: eng
month: '11'
oa: 1
oa_version: Published Version
page: 101-113
pmid: 1
project:
- _id: 34ae1506-11ca-11ed-8bc3-c14f4c474396
grant_number: F8810
name: The highjacking of meiosis for asexual reproduction
publication: Matrix Biology
publication_identifier:
eissn:
- 1569-1802
issn:
- 0945-053X
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
status: public
title: Evidence for the major role of PH4⍺EFB in the prolyl 4-hydroxylation of Drosophila
collagen IV
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: 141
year: '2025'
...
---
_id: '6297'
abstract:
- lang: eng
text: Cell-cell and cell-glycocalyx interactions under flow are important for the
behaviour of circulating cells in blood and lymphatic vessels. However, such interactions
are not well understood due in part to a lack of tools to study them in defined
environments. Here, we develop a versatile in vitro platform for the study of
cell-glycocalyx interactions in well-defined physical and chemical settings under
flow. Our approach is demonstrated with the interaction between hyaluronan (HA,
a key component of the endothelial glycocalyx) and its cell receptor CD44. We
generate HA brushes in situ within a microfluidic device, and demonstrate the
tuning of their physical (thickness and softness) and chemical (density of CD44
binding sites) properties using characterisation with reflection interference
contrast microscopy (RICM) and application of polymer theory. We highlight the
interactions of HA brushes with CD44-displaying beads and cells under flow. Observations
of CD44+ beads on a HA brush with RICM enabled the 3-dimensional trajectories
to be generated, and revealed interactions in the form of stop and go phases with
reduced rolling velocity and reduced distance between the bead and the HA brush,
compared to uncoated beads. Combined RICM and bright-field microscopy of CD44+
AKR1 T-lymphocytes revealed complementary information about the dynamics of cell
rolling and cell morphology, and highlighted the formation of tethers and slings,
as they interacted with a HA brush under flow. This platform can readily incorporate
more complex models of the glycocalyx, and should permit the study of how mechanical
and biochemical factors are orchestrated to enable highly selective blood cell-vessel
wall interactions under flow.
article_processing_charge: No
article_type: original
author:
- first_name: Heather S.
full_name: Davies, Heather S.
last_name: Davies
- first_name: Natalia S.
full_name: Baranova, Natalia S.
id: 38661662-F248-11E8-B48F-1D18A9856A87
last_name: Baranova
orcid: 0000-0002-3086-9124
- first_name: Nouha
full_name: El Amri, Nouha
last_name: El Amri
- first_name: Liliane
full_name: Coche-Guérente, Liliane
last_name: Coche-Guérente
- first_name: Claude
full_name: Verdier, Claude
last_name: Verdier
- first_name: Lionel
full_name: Bureau, Lionel
last_name: Bureau
- first_name: Ralf P.
full_name: Richter, Ralf P.
last_name: Richter
- first_name: Delphine
full_name: Débarre, Delphine
last_name: Débarre
citation:
ama: Davies HS, Baranova NS, El Amri N, et al. An integrated assay to probe endothelial
glycocalyx-blood cell interactions under flow in mechanically and biochemically
well-defined environments. Matrix Biology. 2019;78-79:47-59. doi:10.1016/j.matbio.2018.12.002
apa: Davies, H. S., Baranova, N. S., El Amri, N., Coche-Guérente, L., Verdier, C.,
Bureau, L., … Débarre, D. (2019). An integrated assay to probe endothelial glycocalyx-blood
cell interactions under flow in mechanically and biochemically well-defined environments.
Matrix Biology. Elsevier. https://doi.org/10.1016/j.matbio.2018.12.002
chicago: Davies, Heather S., Natalia S. Baranova, Nouha El Amri, Liliane Coche-Guérente,
Claude Verdier, Lionel Bureau, Ralf P. Richter, and Delphine Débarre. “An Integrated
Assay to Probe Endothelial Glycocalyx-Blood Cell Interactions under Flow in Mechanically
and Biochemically Well-Defined Environments.” Matrix Biology. Elsevier,
2019. https://doi.org/10.1016/j.matbio.2018.12.002.
ieee: H. S. Davies et al., “An integrated assay to probe endothelial glycocalyx-blood
cell interactions under flow in mechanically and biochemically well-defined environments,”
Matrix Biology, vol. 78–79. Elsevier, pp. 47–59, 2019.
ista: Davies HS, Baranova NS, El Amri N, Coche-Guérente L, Verdier C, Bureau L,
Richter RP, Débarre D. 2019. An integrated assay to probe endothelial glycocalyx-blood
cell interactions under flow in mechanically and biochemically well-defined environments.
Matrix Biology. 78–79, 47–59.
mla: Davies, Heather S., et al. “An Integrated Assay to Probe Endothelial Glycocalyx-Blood
Cell Interactions under Flow in Mechanically and Biochemically Well-Defined Environments.”
Matrix Biology, vol. 78–79, Elsevier, 2019, pp. 47–59, doi:10.1016/j.matbio.2018.12.002.
short: H.S. Davies, N.S. Baranova, N. El Amri, L. Coche-Guérente, C. Verdier, L.
Bureau, R.P. Richter, D. Débarre, Matrix Biology 78–79 (2019) 47–59.
date_created: 2019-04-11T20:55:01Z
date_published: 2019-05-01T00:00:00Z
date_updated: 2023-08-25T10:11:28Z
day: '01'
ddc:
- '570'
department:
- _id: MaLo
doi: 10.1016/j.matbio.2018.12.002
external_id:
isi:
- '000468707600005'
file:
- access_level: open_access
checksum: 790878cd78bfc54a147ddcc7c8f286a0
content_type: application/pdf
creator: dernst
date_created: 2020-05-14T09:02:07Z
date_updated: 2020-07-14T12:47:27Z
file_id: '7825'
file_name: 2018_MatrixBiology_Davies.pdf
file_size: 4444339
relation: main_file
file_date_updated: 2020-07-14T12:47:27Z
has_accepted_license: '1'
isi: 1
language:
- iso: eng
month: '05'
oa: 1
oa_version: Submitted Version
page: 47-59
publication: Matrix Biology
publication_identifier:
issn:
- 0945-053X
publication_status: published
publisher: Elsevier
quality_controlled: '1'
status: public
title: An integrated assay to probe endothelial glycocalyx-blood cell interactions
under flow in mechanically and biochemically well-defined environments
tmp:
image: /images/cc_by_nc_nd.png
legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode
name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
(CC BY-NC-ND 4.0)
short: CC BY-NC-ND (4.0)
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 78-79
year: '2019'
...