The molecular basis for substrate recognition and gating in sugar transporters

Sammanfattning: Sugar is a vital sustenance for most forms of life. For a cell to take up sugar, specialized transport proteins embedded into the membrane bilayer known as sugar porters, are required. Dysfunction of sugar porters is associated with some metabolic diseases, and their expression is upregulated in many cancers as they typically require more sugar than normal cells. Furthermore, sugar porters also play a role in the vitality of the malaria parasite.The mechanism of sugar transport is known as a rocker-switch alternating access mechanism. Simplistically, sugar binds between two similar domains on the outside of a sugar transporter and the domains then move around the sugar, so the sugar is exposed to the inside. During this domain movement, protein mass will block the sugar binding site from both outside and inside, forming the occluded state which is essential to ensure no substrate leakage during transport. Despite this relatively simple model of transport, little is known about how different sugar porters display diverse substrate specificity, affinity, and turnover.In the four papers making up this thesis, we structurally characterize missing pieces of the sugar transport cycle, identify how these states are connected with simulations, and assess factors contributing to sugar transport by functional assays. With simulations, we show how sugar catalyzes conformational change by interacting with the occluded state. We demonstrate our functional proteoliposome-based transport assay, which allows us to measure the effect of protein mutations, inhibitors, and lipid influences in sugar recognition and turnover. Characterization of the malaria parasite hexose transporter PfHT1 has allowed us to understand antimalarial inhibitor specificity against this protein which could have implications in combating the disease, as well as pharmacological control of sugar porters in general.