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Millisecond dynamics of an unlabeled amino acid transporter

Author

Listed:
  • Tina R. Matin

    (Weill Cornell Medicine
    Weill Cornell Medicine)

  • George R. Heath

    (Weill Cornell Medicine
    Weill Cornell Medicine)

  • Gerard H. M. Huysmans

    (Weill Cornell Medicine)

  • Olga Boudker

    (Weill Cornell Medicine
    Weill Cornell Medicine)

  • Simon Scheuring

    (Weill Cornell Medicine
    Weill Cornell Medicine)

Abstract

Excitatory amino acid transporters (EAATs) are important in many physiological processes and crucial for the removal of excitatory amino acids from the synaptic cleft. Here, we develop and apply high-speed atomic force microscopy line-scanning (HS-AFM-LS) combined with automated state assignment and transition analysis for the determination of transport dynamics of unlabeled membrane-reconstituted GltPh, a prokaryotic EAAT homologue, with millisecond temporal resolution. We find that GltPh transporters can operate much faster than previously reported, with state dwell-times in the 50 ms range, and report the kinetics of an intermediate transport state with height between the outward- and inward-facing states. Transport domains stochastically probe transmembrane motion, and reversible unsuccessful excursions to the intermediate state occur. The presented approach and analysis methodology are generally applicable to study transporter kinetics at system-relevant temporal resolution.

Suggested Citation

  • Tina R. Matin & George R. Heath & Gerard H. M. Huysmans & Olga Boudker & Simon Scheuring, 2020. "Millisecond dynamics of an unlabeled amino acid transporter," Nature Communications, Nature, vol. 11(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-18811-z
    DOI: 10.1038/s41467-020-18811-z
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    Cited by:

    1. Alma P. Perrino & Atsushi Miyagi & Simon Scheuring, 2021. "Single molecule kinetics of bacteriorhodopsin by HS-AFM," Nature Communications, Nature, vol. 12(1), pages 1-10, December.
    2. Yangang Pan & Jingyu Zhan & Yining Jiang & Di Xia & Simon Scheuring, 2023. "A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1," Nature Communications, Nature, vol. 14(1), pages 1-13, December.

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