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Related Concept Videos

Single-pass Transmembrane Proteins01:25

Single-pass Transmembrane Proteins

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Integral membrane proteins are tightly associated with the cell membrane and play a crucial role in cell communication, signaling, adhesion, and transport of the molecules. Some integral membrane proteins are present only in the membrane monolayer. For example, the enzyme fatty acid amide hydrolase is present in the cytoplasmic side of the membrane monolayer. In contrast, another type of integral membrane protein, also known as a transmembrane protein, spans across the membrane. Transmembrane...
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Insertion of Multi-pass Transmembrane Proteins in the RER01:29

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The rough ER membrane synthesizes, assembles, and embeds transmembrane proteins in diverse topologies. These proteins function as transporters or channels and can remain in the ER membrane or are sent to the Golgi complex, lysosome, and cell membrane.
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Insertion of Single-pass Transmembrane Proteins in the RER01:26

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Integral membrane proteins are proteins adhered to the lipid bilayer of a cell organelle or membrane. They can be of two types: transmembrane integral proteins that span the lipid bilayer and monotopic proteins that are attached to either side of the membrane but do not pass through it.
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Multi-pass Transmembrane Proteins and β-barrels01:09

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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
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Protein Transport into the Inner Mitochondrial Membrane01:34

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Nuclear encoded mitochondrial precursors are imported to the inner membrane in a multistep process involving two separate translocons, TIM22 and TIM23. TIM23 is a cation-selective pore that remains closed by the N terminal segment of the protein. Negative charges on the TIM23 act as a receptor for the incoming precursor, pulling the positively charged matrix-targeting sequence for peptide insertion and translocation.
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Mechanisms of Membrane-bending01:15

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The living membranes are flexible due to their fluid mosaic nature; however, their bending into different shapes is an active process regulated by specific lipids and proteins. The membrane bending can be transient as seen in vesicles or stable for a long time as in microvilli. Cells regulate the size, location, and duration of the membrane curvature.
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Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
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Integral membrane protein structure determination using pseudocontact shifts.

Duncan J Crick1, Jue X Wang, Bim Graham

  • 1Department of Biochemistry, University of Cambridge, Cambridge, UK.

Journal of Biomolecular NMR
|January 22, 2015
PubMed
Summary
This summary is machine-generated.

Lanthanide-induced pseudocontact shifts (PCSs) aid nuclear magnetic resonance (NMR) structure determination for challenging membrane proteins. This method, using PCS restraints, successfully elucidated protein structures when limited nuclear Overhauser effect (NOE) data was insufficient.

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Biophysics

Background:

  • Nuclear magnetic resonance (NMR) structure determination of large proteins, especially membrane proteins, is often limited by insufficient experimental restraints.
  • Extensive deuteration strategies, while improving spectral quality, can reduce the number of obtainable nuclear Overhauser effect (NOE) restraints, necessitating complementary approaches.
  • Lanthanide-induced pseudocontact shifts (PCSs) have emerged as a valuable tool for determining the structures of globular proteins.

Purpose of the Study:

  • To demonstrate the applicability of lanthanide-induced pseudocontact shifts (PCSs) for the structural determination of integral membrane proteins.
  • To evaluate the utility of PCS-derived restraints in overcoming limitations posed by sparse NOE data in membrane protein structure elucidation.
  • To establish PCS as a viable strategy for challenging polytopical alpha-helical membrane proteins.

Main Methods:

  • Utilized lanthanide binding tags attached to four distinct positions on the 7TM alpha-helical microbial receptor pSRII.
  • Collected PCS data from these lanthanide-labeled positions.
  • Combined PCS-derived restraints with a limited set of NOE restraints for structure calculation.
  • Compared the structure determination outcome with and without PCS restraints using the same NOE dataset.

Main Results:

  • PCS-derived restraints, when combined with a limited set of NOEs, successfully facilitated the backbone structure determination of the integral membrane protein pSRII.
  • The same limited set of NOEs alone was insufficient to determine the correct 3D fold of the protein.
  • This highlights the effectiveness of PCS in providing crucial long-range distance and orientation information for membrane proteins.

Conclusions:

  • Lanthanide-induced pseudocontact shifts (PCSs) represent a powerful and complementary strategy for NMR structure determination of integral membrane proteins.
  • The PCS-based approach is particularly beneficial for challenging polytopical alpha-helical membrane proteins where traditional methods often fall short.
  • The ease of measurement makes PCS an attractive technique for advancing the structural understanding of large and complex membrane protein systems.