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Eukaryotic flagella are conserved multifunctional organelles with roles in motility, intercellular interactions, and signal transduction. Leishmania possess a single flagellum at all stages of their life cycle. Flagella of promastigote forms in the fly are long and motile, with a canonical 9 + 2 microtubule axoneme and an extra-axonemal paraflagellar rod (PFR). This protocol describes a simple method for the isolation of Leishmania mexicana promastigote flagella, optimized to yield intact flagella that retain both the cytoskeletal elements (9 + 2 axoneme and PFR) and the surrounding membrane. The isolated flagella and deflagellated cell bodies are suitable for analysis by electron microscopy, protein mass spectrometry, and lipidomics.

Original publication

DOI

10.1007/978-1-0716-0294-2_29

Type

Journal article

Journal

Methods in molecular biology (Clifton, N.J.)

Publication Date

01/2020

Volume

2116

Pages

485 - 495

Addresses

Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.

Keywords

Flagella, Cytoskeleton, Leishmania mexicana, Protozoan Proteins, Microscopy, Electron, Centrifugation, Density Gradient, Cell Fractionation, Life Cycle Stages, Mass Spectrometry, Lipidomics