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In contrast to passive transport, active transport involves a substance being moved through membranes in a direction against its concentration or electrochemical gradient. There are two types of active transport: primary active transport and secondary active transport. Primary active transport utilizes chemical energy from ATP to drive protein pumps embedded in the cell membrane. With energy from ATP, the pumps transport ions against their electrochemical gradients—a direction they would...
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The electron transport chain (ETC) is a crucial metabolic pathway that facilitates energy conversion in prokaryotic and eukaryotic cells. In eukaryotes, the ETC comprises four membrane-associated protein complexes in the inner mitochondrial membrane. In prokaryotes, the ETC in the plasma membrane can vary in composition, with fewer or different complexes depending on the organism and environmental conditions. These complexes transfer electrons from electron donors, such as NADH and FADH2, to...
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ATP-driven pumps, also known as transport ATPases, are integral membrane proteins. They have binding sites for ATP located on the membrane's cytosolic side and the ion-conducting domain in the transmembrane region. These pumps use the free energy released from ATP hydrolysis to move the solutes across cell membranes against an electrochemical gradient.
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V-type pumps are ATP-driven pumps found in the vacuolar membranes of plants, yeast, endosomal and lysosomal membranes of animal cells, plasma membranes of a few specialized eukaryotic cells, and some prokaryotes. They are also known as the V1Vo-ATPase, that couple ATP hydrolysis to transport protons against a concentration gradient.
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Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting
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Conduction pathway for potassium through the E. coli pump KdpFABC.

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Summary
This summary is machine-generated.

Bacteria use the KdpFABC potassium (K+) pump to survive osmotic stress. This study reveals the pump

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Microbiology

Background:

  • Bacteria utilize the KdpFABC complex, an ATP-dependent K+ pump, to maintain intracellular potassium levels during osmotic stress.
  • The KdpFABC complex comprises KdpA (a K+ transporter) and KdpB (a P-type ATPase), with a proposed ion conduction pathway between them.

Purpose of the Study:

  • To elucidate the structural mechanism of the KdpFABC K+ pump, particularly the ion conduction pathway.
  • To investigate the interaction between the KdpA selectivity filter and the KdpB ion binding site.

Main Methods:

  • Reconstitution of the KdpFABC complex into lipid nanodiscs.
  • Cryo-electron microscopy (cryo-EM) to determine the structure of the pump under turnover conditions.
  • ATPase and ion transport assays to validate mutational effects.

Main Results:

  • A 2.1 Å cryo-EM structure of the KdpFABC pump in the E1~P·ADP conformation was obtained.
  • Strong densities indicate K+ ions bound in the KdpA selectivity filter and KdpB binding site.
  • The study identified a hydrophobic tunnel connecting KdpA and KdpB, with water molecules in the vestibule and tunnel, and a low-affinity release site in KdpB.

Conclusions:

  • The KdpFABC pump facilitates K+ transport through a unique tunnel connecting a channel-like KdpA subunit to an ATPase KdpB subunit.
  • The structural and functional data confirm K+ ion passage through the proposed pathway and highlight the pump's mechanism for maintaining essential K+ gradients in bacteria.