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Liposome-Based Study Provides Insight into Cellular Internalization Mechanism of Mosquito-Larvicidal BinAB Toxin

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Abstract

Glycosylphosphatidylinositols (GPIs) anchored proteins are commonly localized onto lipid rafts. These extracellular proteins participate in a variety of cellular functions, including as receptors for viruses and toxins. Intracellular trafficking of World Health Organization recognized mosquito-larvicidal BinAB toxin is mediated via GPI-anchored Cqm1 receptor protein in Culex mosquitoes. We confirmed conformational change in Cqm1 dimer on interaction with BinA/BinB proteins by dynamic light scattering, modelling of hydrodynamic parameters using the atomic structures, and synchrotron Small Angle solution X-ray scattering (SAXS). A reliable model of the receptor–BinB complex was also constructed from joint SAXS/SANS refinement. We confirmed electrostatic interactions of the Cqm1 ectodomain with lipid rafts reconstituted in model membranes and report receptor-dependent impairment of model liposomes by BinA/B proteins. Liposomal disruption was toxin concentration-dependent as monitored by the release of encapsulated carboxyfluorescein dye. Interestingly, BinA alone, without BinB, showed efficient efflux of the fluorescent dye in agreement with the reported high larvicidal activity of BinA variants. The study provides insight into BinA/B toxin internalization mechanism in the membrane model that is toxin internalization is mediated via receptor-dependent pore formation mechanism. It also suggests a tangible and environmentally safe strategy for control of mosquito population.

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Data Availability

Small Angle solution Scattering (SAXS and SANS) data for Cqm1 (SAXS), BinB (SAXS), BinB–Cqm1 complex (SAXS) and BinB–Cqm1 complex (SANS) have been submitted to the Small Angle Scattering Biological Data Bank (SASBDB; https://www.sasbdb.org; Valentini et al. 2015) with submission Ids SASDHR8, SASDHS8, SASDHT8 and SASDHU8, respectively.

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Acknowledgements

We thank Professor R. Chidambaram for encouragement and useful discussions. We also thank Drs. Debasis Sen, Avik Das and Rahul Singh, Bhabha Atomic Research Centre, for the kind help of SAXS data acquisition on Indus-2 synchrotron.

Funding

The research was funded by the Department of Atomic Energy, Government of India.

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Author notes

  1. Mahima Sharma

    Present address: , 13th Floor, 247 Park, Vikhroli West, Mumbai, 400079, India

Authors and Affiliations

  1. Radiation Biology & Health Sciences Division, Bhabha Atomic Research Centre, Mumbai, Maharashtra, 400085, India

    Mahima Sharma, Amit Kumar & Vinay Kumar

  2. Homi Bhabha National Institute, Training School Complex, Anushaktinagar, Mumbai, Maharashtra, 400094, India

    Vinay Kumar

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Contributions

VK conceived and designed the research, analysed data and wrote the manuscript. MS purified the proteins. MS and AK planned and performed liposomal study.

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Correspondence to Vinay Kumar.

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Sharma, M., Kumar, A. & Kumar, V. Liposome-Based Study Provides Insight into Cellular Internalization Mechanism of Mosquito-Larvicidal BinAB Toxin. J Membrane Biol 253, 331–342 (2020). https://doi.org/10.1007/s00232-020-00131-4

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