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Related Concept Videos

Membrane Asymmetry Regulating Transporters01:19

Membrane Asymmetry Regulating Transporters

Enzymes like flippase, floppase, and scramblase transfer phospholipids from one layer to another in the membrane, thereby affecting membrane asymmetry.
Flippase
Eukaryotic flippases are type-IV P-type ATPases or P4-ATPases belonging to P-type ATPase family proteins that are membrane-bound pumps involved in the ATP-mediated transport of ions and molecules across the membrane. Flippases flip specific phospholipids from the outer to the inner leaflet of a membrane. All P4-ATPases have one...
Phosphoinositides and PIPs01:42

Phosphoinositides and PIPs

Phosphoinositides are a group of phospholipids containing a glycerol backbone with two fatty acid chains and a phosphate attached to a myoinositol sugar ring. The inositol head group extends into the cytoplasm, where it is modified by adding phosphate groups to form phosphatidylinositol phosphates or PIPs.
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Multi-pass Transmembrane Proteins and β-barrels01:09

Multi-pass Transmembrane Proteins and β-barrels

In multi-pass transmembrane proteins, the polypeptide chain crosses the membrane more than once. The transmembrane polypeptide chain either forms an α-helix or β-strand structure. α-Helix containing multi-pass transmembrane proteins are ubiquitous, whereas β-strand containing ones are mainly found in gram-negative bacteria, mitochondria, and chloroplasts.
α-Helix containing multi-pass transmembrane proteins
Multi-pass transmembrane proteins such as G-protein-linked receptors (GPCRs) and...
ATP Driven Pumps I: An Overview01:27

ATP Driven Pumps I: An Overview

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.
There are four main types of ATP-driven pumps - P-type, V-type, F-type, and ABC transporter. All these pumps are of varying complexities and are...
Assembly of the Lipid Bilayer in the ER01:28

Assembly of the Lipid Bilayer in the ER

Biological membranes are more than just a barrier separating cell cytoplasm from the outside environment. They are highly dynamic and help maintain the integrity and physiological stability of the cells as well as membrane-bound organelles. Membranes also play vital roles in cell-to-cell and intracellular communication.
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Mechanisms of Membrane Domain Formation00:59

Mechanisms of Membrane Domain Formation

Different physical properties of lipids and proteins allow them to localize and form distinct islands or domains in the membrane. Some membrane domains are formed due to protein-protein interactions, whereas others are formed due to the presence of specific lipids such as sphingolipids and sterols—for example, large proteins, such as bacteriorhodopsin, aggregate and create distinct domains.
Another mechanism for membrane domain formation involves membrane proteins interacting with cytoskeletal...

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PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions
10:58

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions

Published on: July 27, 2017

Mapping functional interactions in a heterodimeric phospholipid pump.

Catheleyne F Puts1, Radhakrishnan Panatala, Hanka Hennrich

  • 1Department of Membrane Enzymology, Bijvoet Center and Institute of Biomembranes, Utrecht University, The Netherlands.

The Journal of Biological Chemistry
|July 14, 2012
PubMed
Summary
This summary is machine-generated.

Type 4 P-type ATPases (P4-ATPases) require Cdc50 proteins for phospholipid transport. Dynamic association between these proteins is crucial for transporter function and generating membrane asymmetry.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Type 4 P-type ATPases (P4-ATPases) are essential for phospholipid transport, establishing membrane asymmetry in cellular compartments.
  • Their functional mechanism has been debated due to structural similarities with cation transporters, questioning if P4-ATPases alone mediate flippase activity.
  • P4-ATPases form obligate heteromeric complexes with Cdc50 proteins, but the nature of their interaction and its role in transport remain unclear.

Purpose of the Study:

  • To elucidate the structural and functional determinants governing the dynamic association between P4-ATPases and Cdc50 proteins.
  • To investigate the role of the Cdc50 ectodomain and specific disulfide bridges in mediating transporter-subunit interactions and catalytic activity.

Main Methods:

  • Utilized domain swapping, site-directed mutagenesis, and random mutagenesis to identify key residues involved in P4-ATPase·Cdc50 heterodimer formation.
  • Employed biochemical and in vivo assays to assess the impact of mutations on subunit binding affinity and P4-ATPase catalytic function.
  • Analyzed conserved disulfide bridges within the Cdc50 ectodomain through functional characterization of cysteine mutants.

Main Results:

  • Identified residues across the Cdc50 subunit that contribute to heterodimer formation with P4-ATPases.
  • Demonstrated that a specific conformation of the Cdc50 ectodomain is critical for selective and functional transporter-subunit interactions.
  • Disruption of conserved disulfide bridges in Cdc50 led to an inverse correlation between subunit binding and P4-ATPase-mediated phospholipid transport.

Conclusions:

  • The dynamic association between P4-ATPases and Cdc50 proteins is essential for the phospholipid transport cycle.
  • The Cdc50 ectodomain's conformation and integrity, particularly its disulfide bridges, play a critical role in regulating this dynamic interaction and overall transporter functionality.
  • These findings provide crucial insights into the mechanism of P4-ATPase-mediated lipid transport and the regulation of membrane asymmetry.