ArticleCellular and molecular bioengineering2019
Dynamics of Non-Canonical Amino Acid-Labeled Intra- and Extracellular Proteins in the Developing Mouse.
Article in Cellular and molecular bioengineering, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 27 citations in OpenAlex.
- Optimizing bio-orthogonal non-canonical amino acid tagging (BONCAT) for low-disruption labeling of Arabidopsis proteins in vivo.Plant physiology · 2026Article
- Extracellular matrix protein composition dynamically changes during murine forelimb development.iScience · 2024Article
- Quantification of local matrix deposition during muscle stem cell activation using engineered hydrogels.bioRxiv : the preprint server for biology · 2024Article
- Intracellular fraction of zona pellucida protein 3 is required for the oocyte-to-embryo transition in mice.Molecular human reproduction · 2023Article
- Unveiling extracellular matrix assembly: Insights and approaches through bioorthogonal chemistry.Materials today. Bio · 2023Review
- An Integrative Biology Approach to Quantify the Biodistribution of AzidohomoalanineCellular and molecular bioengineering · 2023Article
- Mechanical loading is required for initiation of extracellular matrix deposition at the developing murine myotendinous junction.Matrix biology : journal of the International Society for Matrix Biology · 2023Article
- Extracellular matrix dynamics: tracking in biological systems and their implications.Journal of biological engineering · 2022Review
- Mechanisms and Minimization of False Discovery of Metabolic Bioorthogonal Noncanonical Amino Acid Proteomics.Rejuvenation research · 2022Article
- Metabolic labeling of secreted matrix to investigate cell-material interactions in tissue engineering and mechanobiology.Nature protocols · 2022Review
- 3D Mapping Reveals a Complex and Transient Interstitial Matrix During Murine Kidney Development.Journal of the American Society of Nephrology : JASN · 2021Article
- Extracellular matrix at the muscle - tendon interface: functional roles, techniques to explore and implications for regenerative medicine.Connective tissue research · 2021Review
- Comparative Analysis of the Extracellular Matrix Proteome across the Myotendinous Junction.Journal of proteome research · 2020Article
- Perlecan Knockdown Significantly Alters Extracellular Matrix Composition and Organization During Cartilage Development.Molecular & cellular proteomics : MCP · 2020Article
- Using Genetic Code Expansion for Protein Biochemical Studies.Frontiers in bioengineering and biotechnology · 2020Review
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4 authors at 1 institution in 1 country.
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Abstract
introductionMapping protein synthesis and turnover during development will provide insight into functional tissue assembly; however, quantitative
methodsPregnant C57BL/6 mice were injected with Aha or PBS (control) at different embryonic time points. Aha-labeled proteins from homogenized E12.5 and E15.5 embryos were conjugated with diazo biotin-alkyne, bound to NeutrAvidin beads, selectively released, then processed for either SDS-PAGE or LC-MS/MS. For turnover studies, embryos were harvested 0-48 h after Aha injection at E12.5, separated into different cellular fractions based on solubility, and analyzed
resultsWe developed an enhanced method for isolating Aha-labeled proteins from embryos that minimizes background signal from unlabeled proteins and avidin contamination. Approximately 50% of all identified proteins were found only in Aha samples. Comparing proteins present in both Aha and PBS samples, 90% were > 2-fold enriched in Aha-treated embryos. Furthermore, this method could resolve differences in the Aha-labeled proteome between developmental time points. Newly synthesized Aha-labeled proteins were observed by 3 h and peak labeling was around 6 h. Notably, extracellular matrix and cytoskeletal turnover appeared lower than the cytosolic fraction.
conclusionsThe methods developed in this work enable the identification and quantification of protein synthesis and turnover in different tissue fractions during development. This will provide insight into functional tissue assembly and ultimately inform the design of regenerative therapies that seek to promote growth and repair.
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