Biology subjects

Hudson, B. G.

Publications and source records attributed to Hudson, B. G..

4 recordsLinked to original sources

bioRxiv2026

An Alport variant illuminates the bioactivity of the collagen IV α565- α121 scaffold in Bowman's capsule.

Alport syndrome (AS) is a major cause of chronic kidney failure and affects millions of people worldwide. Pathogenic variants in COL4A3, COL4A4, and COL4A5, which encode the collagen IV 345 scaffold, compromise glomerular basement membrane (GBM) structure and function. However, the molecular mechanisms linking the >5,000 reported variants to disease pathology remain poorly understood. To address this gap, we previously examined a distinctive variant, an 8-amino acid "Z-appendage", added to the NC1 domain of the 3 chain. Knock-in mice carrying this variant developed GBM abnormalities and proteinuria, implicating the NC1 hexamer as a critical determinant of GBM function and suggesting that the hexamer surface contains bioactive sites that may mediate signaling and/or organization of macromolecular complexes. Given that approximately 80% of AS cases are associated with COL4A5 variants, including many within the NC1 hexamer, we asked whether relocating the Z-appendage from the 3 NC1 subunit to the 5 subunit produces similar pathology. Strikingly, Col4a5-Z mice did not develop proteinuria and showed only minor changes in GBM morphology. In contrast, the variant induced marked thickening of Bowmans capsule, accompanied by increased deposition of the collagen IV 121 scaffold, increased fibrillar collagen, and cellular deposits. Structural modeling predicts that the collagen IV 565-121 scaffold bearing two Z-appendages adopts an aberrant secondary structure that may stiffen the scaffold and occlude binding sites. Together, these findings reveal a bioactive role for the collagen IV 565-121 scaffold in the Bowmans capsule basement membrane, with potential implications for other 565-121containing tissues such as the aorta and bladder.

molecular biology↗

bioRxiv2026

Collagen IV of basement membrane: V. Bromide-mediated sulfilimine bonds interlock the quaternary structure of NC1-hexamer of scaffolds enabling metazoan evolution.

Collagen-IV (Col-IV) scaffolds, a primordial basement membrane component, enabled animal multicellularity, evolution and adaptation. These scaffolds provide tensile strength and tether macromolecules, forming supramolecular complexes that interact with cell-surface receptors and influence cell-behavior. Triple-helical Col-IV protomers, composed of three -chains, with a trimeric globular NC1-domain at the C-terminus, oligomerize forming a NC1-hexamer structure that connects adjoining protomers of Col-IV121, Col-IV556--121, and Col-IV345 scaffolds. Hexamer formation and stability are driven by the extracellular chloride concentration-"chloride pressure". Hexamer structure is reinforced by six sulfilimine bonds forming covalent crosslinks that weld together trimeric NC1-domains of adjoining protomers. We recently found evidence that sulfilimine bonds, independent of chloride, stabilize the quaternary structure of the Col-IV345 hexamer of the Col-IV345 scaffold. Here, we sought to determine whether this function also pertains to the Col-IV121 scaffold that occurs ubiquitously across the animal kingdom, and whether bromine, a cofactor of peroxidasin in bond formation, are evolutionary conserved. We found that sulfilimine bonds stabilized the quaternary structure of the Col-IV121 hexamer of bovine, mouse and a basal cnidarian, Nematostella vectensis, and that the mechanism of bond formation mediated by peroxidasin and bromide is evolutionary conserved. Analyses of the crystal structure of the NC1-hexamer revealed that sulfilimine bonds covalently fasten a clasp-motif across the trimer-trimer interface, interlocking the domain-swapping region of neighboring subunits, which reinforces the hexamer quaternary structure imposed by chloride conformational constraints. Collectively, our findings reveal that the sulfilimine-bond reinforcement is a critical event in Col-IV scaffold assembly enabling multicellularity, evolution and adaptation of metazoans, beginning with ancient cnidarians.

molecular biology↗

bioRxiv2023

Collagen IV of basement membranes: I. Origin and diversification of COL4 genes enabling animal evolution

Collagen IV is a major component of basement membranes, a specialized form of extracellular matrix that enabled the assembly of multicellular epithelial tissues. In mammals, collagen IV assembles from a family of six -chains (1 to 6), forming three supramolecular scaffolds: Col-IV121, Col-IV345 and Col-IV121-556. The -chains are encoded by six genes (COL4A1-6) that occur in pairs in a head-to-head arrangement. In Alport syndrome, variants in COL4A3, 4 or 5 genes, encoding Col-IV345 scaffold in glomerular basement membrane (GBM), the kidney ultrafilter, cause progressive renal failure in millions of people worldwide. How variants cause dysfunction remains obscure. Here, we gained insights into Col-IV345 function by determining its evolutionary lineage, as revealed from phylogenetic analyses and tissue expression of COL4 gene-pairs. We found that the COL4A[<]1|2[>] gene-pair emerged in basal Ctenophores and Cnidaria phyla and is highly conserved across metazoans. The COL4A[<]1|2[>] duplicated and arose as the progenitor to the COL4A[<]3|4[>] gene-pair in cyclostomes, coinciding with emergence of kidney GBM, and expressed and conserved in jawed-vertebrates, except for amphibians, and a second duplication as the progenitor to the COL4A[<]5|6[>] gene-pair and conserved in jawed-vertebrates. These findings revealed that Col-IV121 is the progenitor scaffold, expressed ubiquitously in metazoan basement membranes, and which evolved into vertebrate Col-IV345 and expressed in GBM. The Col-IV345 scaffold, in comparison, has an increased number of cysteine residues, varying in number with osmolarity of the environment. Cysteines mediate disulfide crosslinks between protomers, an adaptation enabling a compact GBM that withstands the high hydrostatic pressure associated with glomerular ultrafiltration.

genomics↗

bioRxiv2020

Sequence-dependent mechanics of collagen reflect its structural and functional organization

Extracellular matrix mechanics influence diverse cellular functions, yet surprisingly little is known about the mechanical properties of their constituent collagen proteins. In particular, network-forming collagen IV, an integral component of basement membranes, has been far less studied than fibril-forming collagens. A key feature of collagen IV is the presence of interruptions in the triple-helix-defining (Gly-X-Y) sequence along its collagenous domain. Here, we used atomic force microscopy (AFM) to determine the impact of sequence heterogeneity on the local flexibility of collagen IV and of the fibril-forming collagen III. Our extracted flexibility profile of collagen IV reveals that it possesses highly heterogeneous mechanics, ranging from semi-flexible regions as found for fibril-forming collagens to a lengthy region of high flexibility towards its N terminus. A simple model in which flexibility is dictated only by the presence of interruptions fit the extracted profile reasonably well, providing insight into the alignment of chains and demonstrating that interruptions - particularly when coinciding in multiple chains - significantly enhance local flexibility. To a lesser extent, sequence variations within the triple helix lead to variable flexibility, as seen along the continuously triple-helical collagen III. We found this fibril-forming collagen to possess a high-flexibility region around its matrix-metalloprotease (MMP) binding site, suggesting a unique mechanical fingerprint of this region that is key for matrix remodeling. Surprisingly, proline content did not correlate with local flexibility in either collagen type. We also found that physiologically relevant changes in pH and chloride concentration did not alter the flexibility of collagen IV, indicating such environmental changes are unlikely to control its compaction during secretion. Although extracellular chloride ions play a role in triggering collagen IV network formation, they do not appear to modulate the structure of its collagenous domain. Significance StatementCollagens are the predominant proteins in vertebrates, forming diverse hierarchical structures to support cells and form connective tissues. Despite their mechanical importance, surprisingly little is established about the molecular encoding of mechanics. Here, we image single collagen proteins and find that they exhibit variable flexibility along their backbones. By comparing collagens with continuous and discontinuous triple-helix-forming sequences, we find that the type of helix interruption correlates with local flexibility, providing the first steps towards a much-needed map between sequence, structure, and mechanics in these large proteins. Our results inform our understanding of collagens ability to adopt compact conformations during cellular secretion and suggest a physical mechanism by which higher-order structure may be regulated by the distinct molecular properties of different collagens.

biophysics↗