High-speed atomic force microscopy reveals a three-state elevator mechanism in the citrate transporter CitS.

ABSTRACT: The secondary active transporter CitS shuttles citrate across the cytoplasmic membrane of gram-negative bacteria by coupling substrate translocation to the transport of two Na⁺ ions. Static crystal structures suggest an elevator type of transport mechanism with two states: up and down. However, no dynamic measurements have been performed to substantiate this assumption. Here, we use high-speed atomic force microscopy for real-time visualization of the transport cycle at the level of single transporters. Unexpectedly, instead of a bimodal height distribution for the up and down states, the experiments reveal movements between three distinguishable states, with protrusions of ∼0.5 nm, ∼1.0 nm, and ∼1.6 nm above the membrane, respectively. Furthermore, the real-time measurements sh

SUBMITTER: Maity S 

PROVIDER: S-EPMC8833178 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature