The small subunit of Hemilipin2, a new heterodimeric phospholipase A2 from Hemiscorpius lepturus scorpion venom, mediates the antiangiogenic effect of the whole protein

Imen Jridi, Ivana Catacchio, Hafed Majdoub, Delavar Shahbazeddah, Mohamed El Ayeb, Maria Antonia Frassanito, Antonio Solimando, Domenico Ribatti, Angelo Vacca, Lamia Borchani

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Abstract

In a previous study, we reported the identification of Hemilipin, the first secreted heterodimeric phospholipase A2 (sPLA2) from Hemiscorpius lepturus scorpion venom and demonstrated its effective inhibition of all angiogenesis key steps in vitro and in vivo. Here, we aimed to characterize a second sPLA2, Hemilipin2, from the same venom and to elucidate its antiangiogenic effect. The protein was purified by chromatography separation and analyzed by MALDI/TOF mass spectrometry. Its N terminal amino acid sequence was determined by Edman degradation method and the enzymatic activity by fatty acids release assay. Hemilipin2 antiangiogenic activity was investigated by studying its effect in vitro on adhesion, migration and capillary like tube formation of Human Umbilical Vein Endothelial Cells (HUVECs) and Human Pulmonary Artery Endothelial Cells (HPAECs); and in vivo on the chick embryo chorioallantoic membrane (CAM) assay. Data to be presented show that Hemilipin2 is heterodimeric composed by two subunits: the large one has a molecular weight of 12,866 and the small one of 2461 a.m.u. It has a strong calcium-dependent PLA2 activity and impacts angiogenesis in vitro and in vivo without showing any cytotoxic or apoptotic signs. Its chemical modification with p-Bromophenacyl Bromide abolishes the enzymatic activity without affecting the antiangiogenic effect. Furthermore, it has been proved that Hemilipin2 small subunit was able to inhibit blood vessel formation both in vitro and in vivo. These findings may serve as a starting point for the designing of a new generation of specific inhibitor of human angiogenesis at different steps.

Original languageEnglish
Pages (from-to)38-46
Number of pages9
JournalToxicon
Volume126
DOIs
Publication statusPublished - Feb 1 2017

Fingerprint

Secretory Phospholipase A2
Scorpion Venoms
Phospholipases A2
Endothelial cells
Assays
Chorioallantoic Membrane
Angiogenesis Inhibitors
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Human Umbilical Vein Endothelial Cells
Venoms
Chemical modification
Blood vessels
Chick Embryo
Chromatography
Pulmonary Artery
Mass spectrometry
Blood Vessels
Amino Acid Sequence
Mass Spectrometry
Proteins

Keywords

  • Angiogenesis
  • Hemiscorpius lepturus sPLA2 group III
  • HPAEC
  • HUVEC
  • Small subunit

ASJC Scopus subject areas

  • Toxicology

Cite this

The small subunit of Hemilipin2, a new heterodimeric phospholipase A2 from Hemiscorpius lepturus scorpion venom, mediates the antiangiogenic effect of the whole protein. / Jridi, Imen; Catacchio, Ivana; Majdoub, Hafed; Shahbazeddah, Delavar; El Ayeb, Mohamed; Frassanito, Maria Antonia; Solimando, Antonio; Ribatti, Domenico; Vacca, Angelo; Borchani, Lamia.

In: Toxicon, Vol. 126, 01.02.2017, p. 38-46.

Research output: Contribution to journalArticle

Jridi, I, Catacchio, I, Majdoub, H, Shahbazeddah, D, El Ayeb, M, Frassanito, MA, Solimando, A, Ribatti, D, Vacca, A & Borchani, L 2017, 'The small subunit of Hemilipin2, a new heterodimeric phospholipase A2 from Hemiscorpius lepturus scorpion venom, mediates the antiangiogenic effect of the whole protein', Toxicon, vol. 126, pp. 38-46. https://doi.org/10.1016/j.toxicon.2016.12.001
Jridi, Imen ; Catacchio, Ivana ; Majdoub, Hafed ; Shahbazeddah, Delavar ; El Ayeb, Mohamed ; Frassanito, Maria Antonia ; Solimando, Antonio ; Ribatti, Domenico ; Vacca, Angelo ; Borchani, Lamia. / The small subunit of Hemilipin2, a new heterodimeric phospholipase A2 from Hemiscorpius lepturus scorpion venom, mediates the antiangiogenic effect of the whole protein. In: Toxicon. 2017 ; Vol. 126. pp. 38-46.
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abstract = "In a previous study, we reported the identification of Hemilipin, the first secreted heterodimeric phospholipase A2 (sPLA2) from Hemiscorpius lepturus scorpion venom and demonstrated its effective inhibition of all angiogenesis key steps in vitro and in vivo. Here, we aimed to characterize a second sPLA2, Hemilipin2, from the same venom and to elucidate its antiangiogenic effect. The protein was purified by chromatography separation and analyzed by MALDI/TOF mass spectrometry. Its N terminal amino acid sequence was determined by Edman degradation method and the enzymatic activity by fatty acids release assay. Hemilipin2 antiangiogenic activity was investigated by studying its effect in vitro on adhesion, migration and capillary like tube formation of Human Umbilical Vein Endothelial Cells (HUVECs) and Human Pulmonary Artery Endothelial Cells (HPAECs); and in vivo on the chick embryo chorioallantoic membrane (CAM) assay. Data to be presented show that Hemilipin2 is heterodimeric composed by two subunits: the large one has a molecular weight of 12,866 and the small one of 2461 a.m.u. It has a strong calcium-dependent PLA2 activity and impacts angiogenesis in vitro and in vivo without showing any cytotoxic or apoptotic signs. Its chemical modification with p-Bromophenacyl Bromide abolishes the enzymatic activity without affecting the antiangiogenic effect. Furthermore, it has been proved that Hemilipin2 small subunit was able to inhibit blood vessel formation both in vitro and in vivo. These findings may serve as a starting point for the designing of a new generation of specific inhibitor of human angiogenesis at different steps.",
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AU - Majdoub, Hafed

AU - Shahbazeddah, Delavar

AU - El Ayeb, Mohamed

AU - Frassanito, Maria Antonia

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