Deen, M. | bioRxiv Science

bioRxiv2026

A lysyl oxidase (LOX)/bone morphogenetic protein-1 (BMP1) complex to facilitate collagen remodeling

Collagen biosynthesis within the extracellular matrix (ECM) relies on finely regulated enzymatic steps to ensure proper collagen maturation and fibrillar assembly. Among these, bone morphogenetic protein-1 (BMP1) and the canonical lysyl oxidase (LOX) act on the collagen telopeptide to promote procollagen processing and oxidative cross-linking, respectively. However, the mechanisms that ensure precise coordination of their activities remain poorly understood. Using NanoBiT assays, we identified and characterized a stable LOX/BMP1 protein complex that assembles intracellularly during trafficking through the ER/Golgi pathway and persists after secretion. Analysis of BMP1 and LOX domains involved in the interaction showed that BMP1 binding requires its CUB2/3 domains, while LOX recognition depends on a conserved, positively charged segment of LOX (residues 259-285) located immediately upstream of its catalytic domain. Formation of the LOX/BMP1 complex did not substantially alter LOX enzymatic activity but markedly enhanced LOX association with collagen type I through the carboxy-telopeptide region, facilitating the assembly of a ternary LOX/BMP1/collagen complex. This pre-assembled complex promoted efficient targeting of LOX to nascent collagen fibrils. Our findings reveal a previously unrecognized layer of regulation in collagen biosynthesis, in which LOX and BMP1 act as a functional unit to ensure precise localization and proper processing of collagen. This mechanism offers new insights into ECM assembly and identifies the LOX/BMP1 interface as a potentially druggable node for anti-fibrotic strategies.

molecular biology↗

bioRxiv2024

Direct and indirect regulation of β- glucocerebrosidase by the transcription factors USF2 and ONECUT2

Mutations in the GBA gene, which encodes the lysosomal enzyme {beta}-glucocerebrosidase (GCase), are the most prevalent genetic susceptibility factor for Parkinsons disease (PD). However, only approximately 20% of carriers develop the disease, suggesting the presence of genetic modifiers influencing the risk of developing PD in the presence of GBA mutations. Here we screened 1,634 human transcription factors (TFs) for their effect on GCase activity in cell lysates of the human glioblastoma line LN-229, into which we introduced the pathogenic GBA L444P variant via adenine base editing. Using a novel arrayed CRISPR activation library, we uncovered 11 TFs as regulators of GCase activity. Among these, activation of MITF and TFEC increased lysosomal GCase activity in live cells, while activation of ONECUT2 and USF2 decreased it. Conversely, ablating USF2 increased GBA mRNA and led to enhanced levels of GCase protein and activity. While MITF, TFEC, and USF2 affected GBA transcription, ONECUT2 was found to control GCase trafficking by modulating the guanine exchange factors PLEKHG4 and PLEKHG4B. Hence, our study provides a systematic approach to identifying modulators of GCase activity, expands the transcriptional landscape of GBA regulation, and deepens our understanding of the mechanisms involved in influencing GCase activity.

molecular biology↗