jats:titleAbstract</jats:title>jats:pAfrican trypanosomes, such as jats:italicTrypanosoma brucei,</jats:italic> are flagellated protozoa which proliferate in mammals and cause a variety of diseases in people and animals. In a mammalian host, the external face of the African trypanosome plasma membrane is covered by a densely packed coat formed of variant surface glycoprotein (VSG), which counteracts the host's adaptive immune response by antigenic variation. The VSG is attached to the external face of the plasma membrane by covalent attachment of the C-terminus to glycosylphosphatidylinositol. As the trypanosome grows, newly synthesised VSG is added to the plasma membrane by vesicle fusion to the flagellar pocket, the sole location of exo- and endocytosis. Snake venoms contain dozens of components, including proteases and phospholipases Ajats:sub2</jats:sub>. Here, we investigated the effect of jats:italicNaja nigricollis</jats:italic> venom on jats:italicT. brucei</jats:italic> with the aim of describing the response of the trypanosome to hydrolytic attack on the VSG. We found no evidence for VSG hydrolysis, however, jats:italicN. nigricollis</jats:italic> venom caused: (i) an enlargement of the flagellar pocket, (ii) the Rab11 positive endosomal compartments to adopt an abnormal dispersed localisation, and (iii) cell cycle arrest prior to cytokinesis. Our results indicate that a single protein family, the phospholipases Ajats:sub2</jats:sub> present in jats:italicN. nigricollis</jats:italic> venom, may be necessary and sufficient for the effects. This study provides new molecular insight into jats:italicT. brucei</jats:italic> biology and possibly describes mechanisms that could be exploited for jats:italicT. brucei</jats:italic> targeting.</jats:p>