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Related Experiment Videos

Triplin: Mechanistic Basis for Voltage Gating.

Marco Colombini1, Patrick Liu1, Chase Dee1

  • 1Department of Biology, University of Maryland, College Park, MD 20842, USA.

International Journal of Molecular Sciences
|July 29, 2023
PubMed
Summary
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Triplin, a novel bacterial pore-former, exhibits complex, sequential voltage-dependent gating. Its unique three-pore structure closes via a translocating voltage sensor, blocking the pores.

Area of Science:

  • Biophysics
  • Microbiology
  • Structural Biology

Background:

  • Gram-negative bacteria outer membranes feature porins, some with voltage-dependent gating.
  • Existing porins exhibit weak voltage dependence, closing at high potentials.

Purpose of the Study:

  • To investigate the novel bacterial pore-former, Triplin, and elucidate its unique voltage-dependent gating mechanism.
  • To characterize the sequential closure of Triplin's three pores and the role of its voltage sensor.

Main Methods:

  • Utilized kinetic measurements to analyze the pore closure process.
  • Investigated voltage-dependent blockage using polyarginine and polylysine.

Main Results:

  • Triplin possesses a highly voltage-dependent, three-pore structure with sequential gating: pore 1 (positive), pore 2 (negative), and pore 3 (positive potentials).
Keywords:
cooperativitykineticspolyarginineporeporinprokaryotevoltage dependencevoltage sensor

Related Experiment Videos

  • A positively charged domain (voltage sensor) translocates through the membrane, blocking the pores during closure.
  • Polyarginine blocked Triplin pores in a voltage-dependent manner, unlike polylysine, supporting the proposed sensor-binding model.

    • Conclusions:

    • Triplin's complex gating mechanism involves a translocating voltage sensor primarily composed of arginine residues.
    • Each pore contains a complementary binding site for the voltage sensor, facilitating pore blockage.
    • The findings provide novel insights into voltage-dependent pore formation and gating in bacteria.