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Developing giant plasma membrane vesicles from Leishmania cells to investigate the role of membrane proteins in photodynamic inactivation.

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Title: Developing giant plasma membrane vesicles from Leishmania cells to investigate the role of membrane proteins in photodynamic inactivation.
Authors: de Souza, Maressa D. F.1 (AUTHOR), Ciancaglini, Pietro2 (AUTHOR), Itri, Rosangela3 (AUTHOR) itri@if.usp.br, Ribeiro, Martha S.1 (AUTHOR) marthasr@usp.br
Source: Photochemistry & Photobiology. Mar/Apr2026, Vol. 102 Issue 2, p480-488. 9p.
Subjects: Membrane proteins, Photooxidation, Oxidative stress, Membrane permeability (Biology), Leishmania, Phototherapy, Extracellular vesicles, Apoptosis
Abstract: Interest in antimicrobial photodynamic therapy for treating cutaneous leishmaniasis has been rising, showing promising outcomes and good patient tolerance. In this study, we aimed to develop a protocol for producing giant plasma membrane vesicles (GPMVs) from Leishmania amazonensis promastigote cell membranes, focusing on the role of membrane‐embedded proteins during methylene blue (MB) photooxidation with red light. Membrane extraction was achieved via centrifugation with various sucrose gradients. We then generated GPMVs by electroformation, applying different frequencies and voltages over four cycles, and examined them using phase contrast optical microscopy. For MB photooxidation, GPMVs were dispersed in an aqueous solution with 50 μM MB and exposed to 665 nm light at 830 μW. A comparable approach was used for mimetic membranes (giant unilamellar vesicles, GUVs) made of Leishmania membrane lipids. MB photoactivation in GUVs caused a transient increase in membrane area and full permeability. Conversely, GPMVs showed an earlier onset of contrast loss but exhibited less overall contrast reduction and no expansion, indicating that membrane proteins in GPMVs modulate the response to oxidative stress. Real‐time monitoring revealed morphological changes in L. amazonensis promastigote cells consistent with apoptosis following photodynamic inactivation. [ABSTRACT FROM AUTHOR]
Copyright of Photochemistry & Photobiology is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
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DbLabel: Engineering Source
An: 193923673
AccessLevel: 6
PubType: Academic Journal
PubTypeId: academicJournal
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  Label: Title
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  Data: Developing giant plasma membrane vesicles from Leishmania cells to investigate the role of membrane proteins in photodynamic inactivation.
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  Data: <searchLink fieldCode="JN" term="%22Photochemistry+%26+Photobiology%22">Photochemistry & Photobiology</searchLink>. Mar/Apr2026, Vol. 102 Issue 2, p480-488. 9p.
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  Data: <searchLink fieldCode="DE" term="%22Membrane+proteins%22">Membrane proteins</searchLink><br /><searchLink fieldCode="DE" term="%22Photooxidation%22">Photooxidation</searchLink><br /><searchLink fieldCode="DE" term="%22Oxidative+stress%22">Oxidative stress</searchLink><br /><searchLink fieldCode="DE" term="%22Membrane+permeability+%28Biology%29%22">Membrane permeability (Biology)</searchLink><br /><searchLink fieldCode="DE" term="%22Leishmania%22">Leishmania</searchLink><br /><searchLink fieldCode="DE" term="%22Phototherapy%22">Phototherapy</searchLink><br /><searchLink fieldCode="DE" term="%22Extracellular+vesicles%22">Extracellular vesicles</searchLink><br /><searchLink fieldCode="DE" term="%22Apoptosis%22">Apoptosis</searchLink>
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  Label: Abstract
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  Data: Interest in antimicrobial photodynamic therapy for treating cutaneous leishmaniasis has been rising, showing promising outcomes and good patient tolerance. In this study, we aimed to develop a protocol for producing giant plasma membrane vesicles (GPMVs) from Leishmania amazonensis promastigote cell membranes, focusing on the role of membrane‐embedded proteins during methylene blue (MB) photooxidation with red light. Membrane extraction was achieved via centrifugation with various sucrose gradients. We then generated GPMVs by electroformation, applying different frequencies and voltages over four cycles, and examined them using phase contrast optical microscopy. For MB photooxidation, GPMVs were dispersed in an aqueous solution with 50 μM MB and exposed to 665 nm light at 830 μW. A comparable approach was used for mimetic membranes (giant unilamellar vesicles, GUVs) made of Leishmania membrane lipids. MB photoactivation in GUVs caused a transient increase in membrane area and full permeability. Conversely, GPMVs showed an earlier onset of contrast loss but exhibited less overall contrast reduction and no expansion, indicating that membrane proteins in GPMVs modulate the response to oxidative stress. Real‐time monitoring revealed morphological changes in L. amazonensis promastigote cells consistent with apoptosis following photodynamic inactivation. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Photochemistry & Photobiology is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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RecordInfo BibRecord:
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      – Type: doi
        Value: 10.1111/php.70000
    Languages:
      – Code: eng
        Text: English
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      Pagination:
        PageCount: 9
        StartPage: 480
    Subjects:
      – SubjectFull: Membrane proteins
        Type: general
      – SubjectFull: Photooxidation
        Type: general
      – SubjectFull: Oxidative stress
        Type: general
      – SubjectFull: Membrane permeability (Biology)
        Type: general
      – SubjectFull: Leishmania
        Type: general
      – SubjectFull: Phototherapy
        Type: general
      – SubjectFull: Extracellular vesicles
        Type: general
      – SubjectFull: Apoptosis
        Type: general
    Titles:
      – TitleFull: Developing giant plasma membrane vesicles from Leishmania cells to investigate the role of membrane proteins in photodynamic inactivation.
        Type: main
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          Name:
            NameFull: de Souza, Maressa D. F.
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            NameFull: Ciancaglini, Pietro
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            NameFull: Itri, Rosangela
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            NameFull: Ribeiro, Martha S.
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            – D: 01
              M: 03
              Text: Mar/Apr2026
              Type: published
              Y: 2026
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              Value: 102
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            – TitleFull: Photochemistry & Photobiology
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