Cell migration is dependent on a number of physical and chemical parameters of the substrate that influence cellular signaling events as cell surface receptors interact with the substrate. These events can strengthen or loosen the contact of the cell with its environment and need to be orchestrated for efficient motility. A set of tunable substrates was used in combination with quantitative imaging to probe for potentially subtle differences in genetically modified and chemically treated rapidly migrating cells. As model cell, Plasmodium sporozoites were used, the forms of malaria parasites transmitted by the mosquito to the host. Sporozoites lacking a substrate-binding surface protein moved on different surfaces with consistently lower efficiency and were more sensitive to adhesion ligand spacing than wild type sporozoites. Addition of an actin filament stabilizing chemical agent temporarily increased sporozoite motility on soft but not on hard substrates. Defined conditions were found where the chemical completely compensates the reduced migration capacity of the genetically modified parasites. As the onset of motility was delayed for sporozoites on unfavourable gels it is suggested that the parasite can slowly adjust to environmental elasticity, possibly by adapting the interplay between surface adhesins and actin filament dynamics. This demonstrates the utility of tunable substrates to dissect molecular function in cell adhesion and motility.