The dynamic nature of netrin-1 and the structural basis for glycosaminoglycan fragment-induced filament formation

Meier, Markus; Gupta, Monika; Akgül, Serife; McDougall, Matthew; Imhof, Thomas; Nikodemus, Denise; Reuten, Raphael; Moya‐Torres, Aniel; To, Vu; Ferens, Fraser G.; Heide, Fabian; Padilla-Meier, G. Pauline; Kukura, Philipp; Huang, Wenming; Gerisch, Birgit; Mörgelin, Matthias; Poole, Kate; Dolfi, Luca; Koch, Manuel; Stetefeld, Jörg

doi:10.1038/s41467-023-36692-w

Cited by 7 publications

(3 citation statements)

References 105 publications

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“…During the review of our manuscript, two articles were released that complement our findings on the roles of proteoglycans and GAGs in netrin and/or UNC-5 function. Meier et al (57) observed that vertebrate netrins oligomerize with heparin through interactions with their EGF2 domains, similar to our findings with UNC-6, and breaking this interaction in the nematode model results in the distal tip cell migration phenotype as we have shown for UNC-5. Akkermans et al (58) showed that mouse UNC5D can form oligomers with a glypican, but without contacting the HS chain.…”

Section: Discussionsupporting

confidence: 91%

Structural insights into the formation of repulsive netrin guidance complexes

Priest,

Nichols,

Smock

et al. 2024

Sci. Adv.

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Netrins dictate attractive and repulsive responses during axon growth and cell migration, where the presence of the receptor Uncoordinated-5 (UNC-5) on target cells results in repulsion. Here, we showed that UNC-5 is a heparin-binding protein, determined its structure bound to a heparin fragment, and could modulate UNC-5–heparin affinity using a directed evolution platform or structure-based rational design. We demonstrated that UNC-5 and UNC-6/netrin form a large, stable, and rigid complex in the presence of heparin, and heparin and UNC-5 exclude the attractive UNC-40/DCC receptor from binding to UNC-6/netrin to a large extent. Caenorhabditis elegans with a heparin-binding–deficient UNC-5 fail to establish proper gonad morphology due to abrogated cell migration, which relies on repulsive UNC-5 signaling in response to UNC-6. Combining UNC-5 mutations targeting heparin and UNC-6/netrin contacts results in complete cell migration and axon guidance defects. Our findings establish repulsive netrin responses to be mediated through a glycosaminoglycan-regulated macromolecular complex.

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Section: Discussionsupporting

confidence: 91%

Structural insights into the formation of repulsive netrin guidance complexes

Priest,

Nichols,

Smock

et al. 2024

Sci. Adv.

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“…( B ) Fibroblast growth factor 2 (FGF2) in complex with fibroblast growth factor receptor 1 (FGFR1), which is mediated by a long-chain heparin ligand (PDB ID: 1FQ9) [ 25 ]. ( C ) Netrin-1 (Net-1) in complex with sucrose octasulfate (SOS) (PDB ID: 7LRF) [ 32 ]. ( D ) Platelet factor 4 (PF4) in complex with the heparin mimetic fondaparinux (PDB ID: 4R9W) [ 33 ].…”

Section: Figurementioning

confidence: 99%

Heparin Mimetics and Their Impact on Extracellular Matrix Protein Assemblies

2023

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Heparan sulfate is a crucial extracellular matrix component that organizes structural features and functional protein processes. This occurs through the formation of protein–heparan sulfate assemblies around cell surfaces, which allow for the deliberate local and temporal control of cellular signaling. As such, heparin-mimicking drugs can directly affect these processes by competing with naturally occurring heparan sulfate and heparin chains that then disturb protein assemblies and decrease regulatory capacities. The high number of heparan-sulfate-binding proteins that are present in the extracellular matrix can cause obscure pathological effects that should be considered and examined in more detail, especially when developing novel mimetics for clinical use. The objective of this article is to investigate recent studies that present heparan-sulfate-mediated protein assemblies and the impact of heparin mimetics on the assembly and function of these protein complexes.

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“…S1) containing the mutant sequence (GeneArt String, Invitrogen) replaced the respective wild-type sequence and was amplified by PCR. Wild type and mutant PCR products were cloned into a modified pCEP-pu vector containing an N-terminal BM40 signal peptide and an N-terminal One-STrEP-tag or an N-terminal His 8 tag [38,39]. The recombinant plasmids were introduced into HEK293-EBNA cells (Invitrogen) using the FuGENE 6 transfection reagent (Roche Applied Science).…”

Section: Introductionmentioning

confidence: 99%

The UCMD-Causing COL6A1 ( $c . 930 + 189 C > T$ ) Intron Mutation Leads to the Secretion and Aggregation of Single Mutated Collagen VI α1 Chains

Freiburg,

Solomon-Degefa,

Freiburg

et al. 2023

Human Mutation

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Collagen VI is a unique member of the collagen family. Its assembly is a complex multistep process and the vulnerability of the process is manifested in muscular diseases. Mutations in COL6A1, COL6A2, and COL6A3 lead to the severe Ullrich Congenital Muscular Dystrophy (UCMD) and a spectrum of disease of varying severity including the milder Bethlem muscular dystrophy. The recently identified dominant intronic mutation in COL6A1 ( c . 930 + 189 C > T ) leads to the partial in-frame insertion of a pseudoexon between exon 11 and exon 12. The pseudoexon is translated into 24 amino acid residues in the N-terminal region of the triple helix and results in the interruption of the typical G-X-Y motif. This recurrent de novo mutation leads to UCMD with a severe progression within the first decade of life. Here, we demonstrate that a mutation-specific antibody detects the mutant chain colocalizing with wild type collagen VI in the endomysium in patient muscle. Surprisingly, in the cell culture of patient dermal fibroblasts, the mutant chain is secreted as a single α chain, while in parallel, normal collagen VI tetramers are assembled with the wild-type α1 chain. The mutant chain cannot be incorporated into collagen VI tetramers but forms large aggregates in the extracellular matrix that may retain the ability to interact with collagen VI and potentially with other molecules. Also, α1 chain-deficient WI-26 VA4 cells transfected with the mutant α1 chain do not assemble collagen VI tetramers but, instead, form aggregates. Interestingly, both the wild type and the mutant single α1 chains form amorphous aggregates when expressed in HEK293 cells in the absence of α2 and α3 chains. The detection of aggregated, assembly incompetent, mutant collagen VI α1 chains provides novel insights into the disease pathophysiology of UCMD patients with the COL6A1 ( c . 930 + 189 C > T ) mutation.

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The dynamic nature of netrin-1 and the structural basis for glycosaminoglycan fragment-induced filament formation

Cited by 7 publications

References 105 publications

Structural insights into the formation of repulsive netrin guidance complexes

Structural insights into the formation of repulsive netrin guidance complexes

Heparin Mimetics and Their Impact on Extracellular Matrix Protein Assemblies

The UCMD-Causing COL6A1 ( $c . 930 + 189 C > T$ ) Intron Mutation Leads to the Secretion and Aggregation of Single Mutated Collagen VI α1 Chains

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The dynamic nature of netrin-1 and the structural basis for glycosaminoglycan fragment-induced filament formation

Cited by 7 publications

References 105 publications

Structural insights into the formation of repulsive netrin guidance complexes

Structural insights into the formation of repulsive netrin guidance complexes

Heparin Mimetics and Their Impact on Extracellular Matrix Protein Assemblies

The UCMD-Causing COL6A1 ( c . 930 + 189 C > T ) Intron Mutation Leads to the Secretion and Aggregation of Single Mutated Collagen VI α1 Chains

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The UCMD-Causing COL6A1 ( $c . 930 + 189 C > T$ ) Intron Mutation Leads to the Secretion and Aggregation of Single Mutated Collagen VI α1 Chains