Residue numbers refer to hDAT. the dopamine (DA) transporter (DAT)1C3, serotonin transporter (SERT)4C6 and norepinephrine transporter (NET) which have critical roles in regulating neurotransmission at synapses; the SLC6 family also includes the glycine transporter, GABA transporter and neutral amino acid transporter subfamilies, in addition to the MAT subfamily. Dysregulation of MATs has been linked to depression, anxiety disorder, attention-deficit-hyperactivity disorder, obsessive-compulsive disorder, substance-use disorders7, epilepsy, Parkinsons disease and autism-spectrum disorder8C11; thus, MATs serve as pharmacological targets for several neuropsychiatric and neurodegenerative disorders. The surge in structure-based studies of MATs in recent years has been driven by the resolution of structures for DAT and SERT. Structural data now permit elucidation of the specific modes of actions of these transporters, beyond those originally deduced from their bacterial homologs, such as the leucine transporter LeuT. LeuT has long served as a prototype for mechanistic studies of NSS function, being the first structurally resolved member of the family12C14. However, tens of structures have now been resolved for DAT (dDAT) and human SERT (hSERT) in multiple conformational states and in the presence of substrate and ions, as well as in the presence of antidepressants and/or psychostimulants. These provide an excellent starting point for delineating the functional dynamics of MATs and their modulation by ligands or drugs, as molecular dynamics (MD) simulations suggest15. The resolution of MAT structures is an extremely important milestone, as well as a solid basis for leveraging advances in computational and experimental technologies to gain a deeper understanding of their mechanisms of function Hpse and interactions. Interactions with small molecules and ions serve a major role in mediating MAT function. First, these are, by definition, secondary transporters; i.e., the transport of their substrate (DA, serotonin, norepinephrine or other monoamines) is effectuated by co-transport of Na+ ions down their electrochemical gradient from the extracellular (EC) medium to the intracellular (IC) medium. Second, eukaryotic members of the SLC6 family, including MATs, are chloride dependent. Notably, even the binding events of the neurotransmitter itself, in addition to those of Na+ and Cl? ions, elicit cooperative changes in the conformation of the transmembrane (TM) domain, which facilitate its translocation. Third, some SLC6 members couple substrate influx to the reverse transport (efflux) of a K+ ion16. Malathion Additionally, the immediate environment has a role; for example, influx Malathion of IC water after the disruption Malathion of IC gates facilitates the dislocation of the substrate from its binding pocket and its release17, and cholesterol (CHOL) molecules affect the transporters conformational dynamics18C21. Most importantly, many small molecules, inhibitors and modulators, including drugs of abuse such as cocaine and amphetamine (AMPH), affect the structural dynamics of MATs. In addition to interactions with small molecules, interactions Malathion with other proteins, or even self interactions to form assemblies or oligomers, enable, enhance or alter transport activity. Many studies have indicated the ability of MATs to co-exist as oligomers22C26 and the effect of intermolecular interactions on reversing transport10,11,27. For example, DA efflux is induced by AMPH28,29 and is regulated by G protein – and -subunits30, the kinase CaMKII28,31, protein kinase C32 and/or phosphatidylinositol-4,5-bisphosphate (PIP2)33,34. Our goal is to provide here an overview of the current understanding of the molecular basis of the mechanisms.