DiDBiT-TMT: A Novel Method to Quantify Changes in the Proteomic Landscape Induced by Neural Plasticity.

Autor: Gamaleldin M; Danish Research Institute of Translational Neuroscience DANDRITE-Nordic EMBL Partnership for Molecular Medicine, Aarhus University, Aarhus C 8000, Denmark.; Department of Molecular Biology and Genetics, Aarhus C 8000, Denmark.; School of Biotechnology, Nile University, Giza 12588, Egypt., Yu NK; Departments of Molecular Medicine and Neurobiology, the Scripps Research Institute, La Jolla 92037, California, United States., Diedrich JK; Departments of Molecular Medicine and Neurobiology, the Scripps Research Institute, La Jolla 92037, California, United States., Ma Y; Departments of Molecular Medicine and Neurobiology, the Scripps Research Institute, La Jolla 92037, California, United States., Wienand A; The Danish National Research Foundation Center of Excellence PROMEMO, Aarhus University, Aarhus C 8000, Denmark.; Danish Research Institute of Translational Neuroscience DANDRITE-Nordic EMBL Partnership for Molecular Medicine, Aarhus University, Aarhus C 8000, Denmark.; Department of Molecular Biology and Genetics, Aarhus C 8000, Denmark., McClatchy DB; Departments of Molecular Medicine and Neurobiology, the Scripps Research Institute, La Jolla 92037, California, United States., Nykjaer A; The Danish National Research Foundation Center of Excellence PROMEMO, Aarhus University, Aarhus C 8000, Denmark.; Danish Research Institute of Translational Neuroscience DANDRITE-Nordic EMBL Partnership for Molecular Medicine, Aarhus University, Aarhus C 8000, Denmark.; Department of Biomedicine, Aarhus University, Aarhus C 8000, Denmark., Nabavi S; The Danish National Research Foundation Center of Excellence PROMEMO, Aarhus University, Aarhus C 8000, Denmark.; Danish Research Institute of Translational Neuroscience DANDRITE-Nordic EMBL Partnership for Molecular Medicine, Aarhus University, Aarhus C 8000, Denmark.; Department of Molecular Biology and Genetics, Aarhus C 8000, Denmark., Yates JR 3rd; Departments of Molecular Medicine and Neurobiology, the Scripps Research Institute, La Jolla 92037, California, United States.
Jazyk: angličtina
Zdroj: Journal of proteome research [J Proteome Res] 2024 Nov 01; Vol. 23 (11), pp. 4878-4895. Date of Electronic Publication: 2024 Oct 07.
DOI: 10.1021/acs.jproteome.4c00180
Abstrakt: Direct detection of biotinylated proteins (DiDBiT) is a proteomic method that can enrich and detect newly synthesized proteins (NSPs) labeled with bio-orthogonal amino acids with 20-fold improved detectability compared to conventional methods. However, DiDBiT has currently been used to compare only two conditions per experiment. Here, we present DiDBiT-TMT, a method that can be used to quantify NSPs across many conditions and replicates in the same experiment by combining isobaric tandem mass tagging (TMT) with DiDBiT. We applied DiDBiT-TMT to brain slices to determine changes in the de novo proteome that occur after inducing chemical long-term potentiation (cLTP) or treatment with the neuromodulator norepinephrine. We successfully demonstrated DiDBiT-TMT's capacity to quantitatively compare up to 9 samples in parallel. We showed that there is a minimal overlap among NSPs that are differentially expressed in cLTP-treated organotypic brain slices, norepinephrine-treated organotypic brain slices, and organotypic slices undergoing combinatorial treatment with norepinephrine and cLTP. Our results point to the possible divergence of the molecular mechanisms underlying these treatments and showcase the applicability of DiDBiT-TMT for studying neurobiology.
Databáze: MEDLINE
Externí odkaz: