Specificity of Loxosceles α clade phospholipase D enzymes for choline-containing lipids: Role of a conserved aromatic cage.

Autor: Moutoussamy EE; Computational Biology Unit, Department of Informatics, University of Bergen, Bergen, Norway.; Department of Biological Sciences, University of Bergen, Bergen, Norway., Waheed Q; Computational Biology Unit, Department of Informatics, University of Bergen, Bergen, Norway.; Department of Biological Sciences, University of Bergen, Bergen, Norway., Binford GJ; Department of Biology, Lewis and Clark College, Portland, Oregon, United States., Khan HM; Computational Biology Unit, Department of Informatics, University of Bergen, Bergen, Norway.; Department of Biological Sciences, University of Bergen, Bergen, Norway., Moran SM; Department of Chemistry and Biochemistry, University of Arizona, Arizona, United States., Eitel AR; Department of Chemistry and Biochemistry, University of Arizona, Arizona, United States., Cordes MHJ; Department of Chemistry and Biochemistry, University of Arizona, Arizona, United States., Reuter N; Computational Biology Unit, Department of Informatics, University of Bergen, Bergen, Norway.; Department of Chemistry, University of Bergen, Bergen, Norway.
Jazyk: angličtina
Zdroj: PLoS computational biology [PLoS Comput Biol] 2022 Feb 18; Vol. 18 (2), pp. e1009871. Date of Electronic Publication: 2022 Feb 18 (Print Publication: 2022).
DOI: 10.1371/journal.pcbi.1009871
Abstrakt: Spider venom GDPD-like phospholipases D (SicTox) have been identified to be one of the major toxins in recluse spider venom. They are divided into two major clades: the α clade and the β clade. Most α clade toxins present high activity against lipids with choline head groups such as sphingomyelin, while activities in β clade toxins vary and include preference for substrates containing ethanolamine headgroups (Sicarius terrosus, St_βIB1). A structural comparison of available structures of phospholipases D (PLDs) reveals a conserved aromatic cage in the α clade. To test the potential influence of the aromatic cage on membrane-lipid specificity we performed molecular dynamics (MD) simulations of the binding of several PLDs onto lipid bilayers containing choline headgroups; two SicTox from the α clade, Loxosceles intermedia αIA1 (Li_αIA) and Loxosceles laeta αIII1 (Ll_αIII1), and one from the β clade, St_βIB1. The simulation results reveal that the aromatic cage captures a choline-headgroup and suggest that the cage plays a major role in lipid specificity. We also simulated an engineered St_βIB1, where we introduced the aromatic cage, and this led to binding with choline-containing lipids. Moreover, a multiple sequence alignment revealed the conservation of the aromatic cage among the α clade PLDs. Here, we confirmed that the i-face of α and β clade PLDs is involved in their binding to choline and ethanolamine-containing bilayers, respectively. Furthermore, our results suggest a major role in choline lipid recognition of the aromatic cage of the α clade PLDs. The MD simulation results are supported by in vitro liposome binding assay experiments.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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