The Role of Intraorganellar Ca2+In Late Endosome–Lysosome Heterotypic Fusion and in the Reformation of Lysosomes from Hybrid Organelles
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We have investigated the requirement for Ca2+ in the fusion and content mixing of rat hepatocyte late endosomes and lysosomes in a cell-free system. Fusion to form hybrid organelles was inhibited by 1,2-bis(2-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid (BAPTA), but not by EGTA, and this inhibition was reversed by adding additional Ca2+. Fusion was also inhibited by methyl ester of EGTA (EGTA-AM), a membrane permeable, hydrolyzable ester of EGTA, and pretreatment of organelles with EGTA-AM showed that the chelation of lumenal Ca2+ reduced the amount of fusion. The requirement for Ca2+ for fusion was a later event than the requirement for a rab protein since the system became resistant to inhibition by GDP dissociation inhibitor at earlier times than it became resistant to BAPTA. We have developed a cell-free assay to study the reformation of lysosomes from late endosome–lysosome hybrid organelles that were isolated from the rat liver. The recovery of electron dense lysosomes was shown to require ATP and was inhibited by bafilomycin and EGTA-AM. The data support a model in which endocytosed Ca2+ plays a role in the fusion of late endosomes and lysosomes, the reformation of lysosomes, and the dynamic equilibrium of organelles in the late endocytic pathway.
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Endosomal membrane fusion events are Ca2+-mediated and likely modulated by changing local Ca2+ gradients.
“Endosomal membrane fusion events, which are essential processes in melanosome and lysosome-related organelle biogenesis, are Ca 2ϩ -mediated (49, 50) and are likely modulated by changing local Ca 2ϩ gradients.”
From SLC24A5 encodes a trans-Golgi network protein with potassium-dependent sodium-calcium exchange activity that regulates human epidermal melanogenesis (2008) · cites this paper as [50]
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- Authors
- Pryor, P. R., Mullock, B. M., Bright, N. A., Gray, S. R., & Luzio, J. P.
- Affiliations
- University of Cambridge (GB)
- Venue
- The Journal of Cell Biology
- Publisher
- Rockefeller University Press
- Published
- 2000-05-29
- Volume
- 149
- Issue
- 5
- Pages
- 1053-1062
- ISSN
- 0021-9525 (ISSN-L)
- 1540-8140
- PMC
- PMC2174832
- OpenAlex
- W2073679788
- Cited by
- 347
- References
- 61
- Citations by year
- 2026: 2 · 2025: 12 · 2024: 7 · 2023: 9 · 2022: 10 · 2021: 20 · 2020: 12 · 2019: 12 · 2018: 9 · 2017: 18 · 2016: 20 · 2015: 20 · 2014: 16 · 2013: 12 · 2012: 21
- FWCI
- 5.53
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- cc-by-nc-sa
- Locations
- The Journal of Cell Biology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
- PubMed Central (publishedVersion, cc-by-nc-sa)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Calcium signaling and nucleotide metabolism
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Keywords
- heterotypic fusion, BAPTA, Rab GTPases, GDP dissociation inhibitor, bafilomycin, ATP, hepatocytes, cell-free assay
- MeSH terms
- Adenosine Triphosphate, Animals, Anti-Bacterial Agents, Calcium, Calmodulin, Cell-Free System, Chelating Agents, Cytosol, Dose-Response Relationship, Drug, Egtazic Acid, Enzyme Inhibitors, Immunoglobulin A, Iodine Radioisotopes, Liver, Lysosomes, Membrane Fusion, Microscopy, Electron, Endosomes, Macrolides, Guanine Nucleotide Dissociation Inhibitors, Rats
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