Abstract
Dysregulation of dopaminergic system induced by HIV-1 Tat protein-mediated direct inhibition of the dopamine transporter (DAT) has been implicated as a mediating factor of HIV-1 associated neurocognitive disorders. We have reported that single point mutations on human DAT (hDAT) at tyrosine88 (Y88F), lysine92 (K92M), and histidine547 (H547A) differentially regulate basal dopamine uptake but diminish Tat-induced inhibition of dopamine uptake by changing dopamine transport process. This study evaluated the effects of double (Y88F/H547A) and triple (Y88F/K92M/H547A) mutations on basal dopamine uptake, Tat-induced inhibition of DAT function, and dynamic transport process. Compared to wild-type hDAT, the Vmax values of [3H]Dopamine uptake were increased by 96% in Y88F/H547A but decreased by 97% in Y88F/K92M/H547A. [3H]WIN35,428 binding sites were not altered in Y88F/H547A but decreased in Y88F/K92M/H547A. Y88F/H547A mutant attenuated Tat-induced inhibition of dopamine uptake observed in wild-type hDAT. Y88F/H547A displayed an attenuation of zinc-augmented [3H]WIN35,428 binding, increased basal dopamine efflux, and reduced amphetamine-induced dopamine efflux, indicating this mutant alters transporter conformational transitions. These findings further demonstrate that both tyrosine88 and histidine547 on hDAT play a key role in stabilizing basal dopamine transport and Tat-DAT integration. This study provides mechanistic insights into developing small molecules to block multiple sites in DAT for Tat binding.
Document Type
Article
Publication Date
3-7-2019
Digital Object Identifier (DOI)
https://doi.org/10.1038/s41598-019-39872-1
Funding Information
Research reported in this publication was supported by National Institutes of Health grants to JZ (R01DA035714 and R21DA041932).
Repository Citation
Sun, Wei-Lun; Quizon, Pamela M.; Yuan, Yaxia; Strauss, Matthew J.; McCain, Richard Jr.; Zhan, Chang-Guo; and Zhu, Jun, "Mutational Effects of Human Dopamine Transporter at Tyrosine88, Lysine92, and Histidine547 on Basal and HIV-1 Tat-Inhibited Dopamine Transport" (2019). Molecular Modeling and Biopharmaceutical Center Faculty Publications. 19.
https://uknowledge.uky.edu/mmbc_facpub/19
Notes/Citation Information
Published in Scientific Reports, v. 9, article no. 3843, p. 1-13.
© The Author(s) 2019
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