دورية أكاديمية
أعرض في EDS

Improved synthesis and application of an alkyne-functionalized isoprenoid analogue to study the prenylomes of motor neurons, astrocytes and their stem cell progenitors.

التفاصيل البيبلوغرافية
العنوان: Improved synthesis and application of an alkyne-functionalized isoprenoid analogue to study the prenylomes of motor neurons, astrocytes and their stem cell progenitors.
المؤلفون: Suazo KF; Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: suazo003@umn.edu., Mishra V; Center for Motor Neuron Biology and Diseases, Department of Neurology. Columbia University Irving Medical Center. New York, NY 10032, USA; Department of Pathology & Cell Biology. Columbia University Irving Medical Center. New York, NY 10032, USA. Electronic address: vartikarmishra@gmail.com., Maity S; Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA., Auger SA; Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: auger054@umn.edu., Justyna K; Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: kjustyna@umn.edu., Petre AM; Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: petre096@umn.edu., Ottoboni L; Department of Pathophysiology and Transplantation, Dino Ferrari Center, Università degli Studi di Milano, Milan, Italy. Electronic address: linda.ottoboni@unimi.it., Ongaro J; Neurology Unit, Foundation IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy., Corti SP; Department of Pathophysiology and Transplantation, Dino Ferrari Center, Università degli Studi di Milano, Milan, Italy; Neurology Unit, Foundation IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy; Neuromuscular and Rare Diseases Unit, Department of Neuroscience, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy. Electronic address: stefania.corti@unimi.it., Lotti F; Center for Motor Neuron Biology and Diseases, Department of Neurology. Columbia University Irving Medical Center. New York, NY 10032, USA; Department of Pathology & Cell Biology. Columbia University Irving Medical Center. New York, NY 10032, USA. Electronic address: fl2219@cumc.columbia.edu., Przedborski S; Center for Motor Neuron Biology and Diseases, Department of Neurology. Columbia University Irving Medical Center. New York, NY 10032, USA; Department of Pathology & Cell Biology. Columbia University Irving Medical Center. New York, NY 10032, USA; Department of Neuroscience, Pathology, and Cell Biology, Columbia University Irving Medical Center, New York, NY 10032, USA. Electronic address: sp30@cumc.columbia.edu., Distefano MD; Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: diste001@umn.edu.
المصدر: Bioorganic chemistry [Bioorg Chem] 2024 Jun; Vol. 147, pp. 107365. Date of Electronic Publication: 2024 Apr 16.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Elsevier Country of Publication: United States NLM ID: 1303703 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1090-2120 (Electronic) Linking ISSN: 00452068 NLM ISO Abbreviation: Bioorg Chem Subsets: MEDLINE
أسماء مطبوعة: Publication: Amsterdam : Elsevier
Original Publication: New York, London, Academic Press.
مواضيع طبية MeSH: Astrocytes*/metabolism , Astrocytes*/cytology , Protein Prenylation* , Alkynes*/chemistry , Alkynes*/chemical synthesis , Motor Neurons*/metabolism , Motor Neurons*/cytology, Animals ; Terpenes/chemistry ; Terpenes/chemical synthesis ; Terpenes/metabolism ; Mice ; Molecular Structure ; Cells, Cultured
مستخلص: Protein prenylation is one example of a broad class of post-translational modifications where proteins are covalently linked to various hydrophobic moieties. To globally identify and monitor levels of all prenylated proteins in a cell simultaneously, our laboratory and others have developed chemical proteomic approaches that rely on the metabolic incorporation of isoprenoid analogues bearing bio-orthogonal functionality followed by enrichment and subsequent quantitative proteomic analysis. Here, several improvements in the synthesis of the alkyne-containing isoprenoid analogue C15AlkOPP are reported to improve synthetic efficiency. Next, metabolic labeling with C15AlkOPP was optimized to obtain useful levels of metabolic incorporation of the probe in several types of primary cells. Those conditions were then used to study the prenylomes of motor neurons (ES-MNs), astrocytes (ES-As), and their embryonic stem cell progenitors (ESCs), which allowed for the identification of 54 prenylated proteins from ESCs, 50 from ES-MNs, and 84 from ES-As, representing all types of prenylation. Bioinformatic analysis revealed specific enriched pathways, including nervous system development, chemokine signaling, Rho GTPase signaling, and adhesion. Hierarchical clustering showed that most enriched pathways in all three cell types are related to GTPase activity and vesicular transport. In contrast, STRING analysis showed significant interactions in two populations that appear to be cell type dependent. The data provided herein demonstrates that robust incorporation of C15AlkOPP can be obtained in ES-MNs and related primary cells purified via magnetic-activated cell sorting allowing the identification and quantification of numerous prenylated proteins. These results suggest that metabolic labeling with C15AlkOPP should be an effective approach for investigating the role of prenylated proteins in primary cells in both normal cells and disease pathologies, including ALS.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier Inc. All rights reserved.)
التعليقات: Update of: bioRxiv. 2024 Mar 06;:. (PMID: 38496415)
معلومات مُعتمدة: R21 NS101575 United States NS NINDS NIH HHS
فهرسة مساهمة: Keywords: Click chemistry; Farnesylation; Geranylgeranylation; Metabolic labeling; Prenylation; Proteomics; Synthetic probe
تواريخ الأحداث: Date Created: 20240418 Date Completed: 20240516 Latest Revision: 20240516
رمز التحديث: 20240517
DOI: 10.1016/j.bioorg.2024.107365
PMID: 38636436
قاعدة البيانات: MEDLINE
الوصف
تدمد:1090-2120
DOI:10.1016/j.bioorg.2024.107365