Palmitoylation is a post-translational modification of Alix regulating the membrane organization of exosome-like small extracellular vesicles

Romancino, Daniele P.; Buffa, Valentina; Caruso, Stefano; Ferrara, Ines; Raccosta, Samuele; Notaro, Antonietta; Campos, Yvan; Noto, Rosina; Martorana, Vincenzo; Cupane, Antonio; Giallongo, Agata; d’Azzo, Alessandra; Manno, Mauro; Bongiovanni, Antonella

doi:10.1016/j.bbagen.2018.09.004

Cited by 50 publications

(47 citation statements)

References 67 publications

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“…These data confirmed that OMVs size did not vary in the temperature range investigated and no membrane damages were reported. However, the smooth peak centered at Q ≈ 1 nm −1 (inset of Figure 8a) is related to the thickness of the membrane and has been already detected by SAXS on Alix-positive exosome-like small extracellular vesicles (Romancino et al, 2018). This peak significantly changed the shape and moved towards higher Q values at increasing temperatures, although this variation was modest.…”

Section: Small Angle X-ray Scattering Experimentsmentioning

confidence: 59%

Biophysical Characterization of Membrane Phase Transition Profiles for the Discrimination of Outer Membrane Vesicles (OMVs) From Escherichia coli Grown at Different Temperatures

et al. 2020

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Dynamic Light Scattering (DLS), Small Angle X-ray Scattering (SAXS) and Transmission Electron Microscopy (TEM) are physical techniques widely employed to characterize the morphology and the structure of vesicles such as liposomes or human extracellular vesicles (exosomes). Bacterial extracellular vesicles are similar in size to human exosomes, although their function and membrane properties have not been elucidated in such detail as in the case of exosomes. Here, we applied the above cited techniques, in synergy with the thermodynamic characterization of the vesicles lipid membrane using a turbidimetric technique to the study of vesicles produced by Gram-negative bacteria (Outer Membrane Vesicles, OMVs) grown at different temperatures. This study demonstrated that our combined approach is useful to discriminate vesicles of different origin or coming from bacteria cultured under different experimental conditions. We envisage that in a near future the techniques employed in our work will be further implemented to discriminate complex mixtures of bacterial vesicles, thus showing great promises for biomedical or diagnostic applications.

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Section: Small Angle X-ray Scattering Experimentsmentioning

confidence: 59%

Biophysical Characterization of Membrane Phase Transition Profiles for the Discrimination of Outer Membrane Vesicles (OMVs) From Escherichia coli Grown at Different Temperatures

et al. 2020

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“…The functions of EVs are strictly correlated to the cell phenotype . As a consequence, different applications require EVs from different cells and, therefore, the field of EVs cannot rely on a single cell line.…”

Section: Methods For Improvement Of Upstream Ev Yieldmentioning

confidence: 99%

Scalable Production and Isolation of Extracellular Vesicles: Available Sources and Lessons from Current Industrial Bioprocesses

Paganini

Palmiero

Pócsfalvi

et al. 2019

Biotechnology Journal

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100

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Potential applications of extracellular vesicles (EVs) are attracting increasing interest in the fields of medicine, cosmetics, and nutrition. However, the manufacturing of EVs is currently characterized by low yields. This limitation severely hampers progress in research at the laboratory and clinical scales, as well as the realization of successful and cost-effective EV-based products. Moreover, the high level of heterogeneity of EVs further complicates reproducible manufacturing on a large scale. In this review, possible directions toward the scalable production of EVs are discussed. In particular, two strategies are considered: i) the optimization of upstream unit operations and ii) the exploitation of well-established and mature technologies already in use in other industrial bioprocesses.

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“…Despite these findings, EV proteomics has proven to be challenging due to co-isolation of non-EV proteins such as lipoproteins during EV isolation, which can mask the presence of less abundant, but potentially physiologically important, EV proteins. Palmitoylation can target proteins to the membrane of EVs 22 , suggesting that palmitoylated proteins are enriched in EVs. However, a recent study that analyzed PTMs of exosomes in depth 23 was conducted using a conventional proteomics procedure, and in that case palmitoylation was not detectable because the thioester bonds were cleaved off during the sample preparation and the palmitoyl groups were lost.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive palmitoyl-proteomic analysis identifies distinct protein signatures for large and small cancer-derived extracellular vesicles

Mariscal

Vagner

Kim

et al. 2019

Preprint

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Extracellular vesicles (EVs) are membrane-enclosed particles that play an important role in cancer progression and have emerged as a promising source of circulating biomarkers. Protein S-acylation, also known as palmitoylation, has been proposed as a post-translational mechanism that modulates the dynamics of EV biogenesis and protein cargo sorting. However, technical challenges have limited large-scale profiling of the whole palmitoyl-proteins of EVs. We successfully employed a novel approach that combines low-background acyl-biotinyl exchange (LB-ABE) with label-free proteomics to analyze the palmitoyl proteome of large EVs (L-EVs) and small EVs (S-EVs) from prostate cancer cells. Here we report the first palmitoyl-protein signature of EVs, and demonstrate that L-and S-EVs harbor proteins associated with distinct biological processes and subcellular origin. We identified STEAP1, STEAP2, and ABBC4 as prostate cancer-specific palmitoyl proteins enriched in both EV populations in comparison with the originating cell lines. Importantly, the presence of the above proteins in EVs was significantly reduced upon inhibition of palmitoylation in the producing cells. These results suggest that palmitoylation may be involved in the differential sorting of proteins to distinct EV populations and allow for better detection of disease biomarkers.

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Palmitoylation is a post-translational modification of Alix regulating the membrane organization of exosome-like small extracellular vesicles

Abstract: Beyond its biological relevance, our study also provides the means for a comprehensive structural characterization of EVs.

Cited by 50 publications

References 67 publications

Biophysical Characterization of Membrane Phase Transition Profiles for the Discrimination of Outer Membrane Vesicles (OMVs) From Escherichia coli Grown at Different Temperatures

Biophysical Characterization of Membrane Phase Transition Profiles for the Discrimination of Outer Membrane Vesicles (OMVs) From Escherichia coli Grown at Different Temperatures

Scalable Production and Isolation of Extracellular Vesicles: Available Sources and Lessons from Current Industrial Bioprocesses

Comprehensive palmitoyl-proteomic analysis identifies distinct protein signatures for large and small cancer-derived extracellular vesicles

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