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Conservation of Protein Domains Over Different Proteins02:26

Conservation of Protein Domains Over Different Proteins

Protein domains are small structurally independent units that are part of a single amino acid chain.  Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to form...
Lipids as Anchors01:32

Lipids as Anchors

In the plasma membrane, the lipids forming the bilayer can also act as an anchor to tether proteins to the membrane. The three main types of lipid anchors found in eukaryotes are – prenyl groups, fatty acyl groups, and glycosylphosphatidylinositol or GPI groups. Prenyl and fatty acyl groups act as anchors on the cytosolic surface of the membrane, whereas GPI anchors proteins on the extracellular side.
The carboxy-terminal of most of the prenylated proteins, such as Ras proteins, contains the...
Membrane Domains01:18

Membrane Domains

The membrane domains concentrate specific lipids and proteins at one place within the membrane, which helps in cell signaling, adhesion, and other critical cellular processes. These domains can differ in size, composition, function, and lifespan.
Protein Domains
The membrane comprises a group of distinct proteins responsible for carrying out a cell's specific function. For example, the plasma membrane of the human sperm, or a single germ cell, contains a unique set of proteins in the anterior...
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...
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
IP3/DAG Signaling Pathway01:11

IP3/DAG Signaling Pathway

Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the  phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and produces two-second...

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Related Experiment Video

Updated: Jul 13, 2026

Transmembrane Domain Oligomerization Propensity determined by ToxR Assay
06:45

Transmembrane Domain Oligomerization Propensity determined by ToxR Assay

Published on: May 26, 2011

Strengthened arm-dimerization domain interactions in AraC.

M Wu1, R Schleif

  • 1Biology Department, Johns Hopkins University, Baltimore, Maryland 21218, USA.

The Journal of Biological Chemistry
|November 9, 2000
PubMed
Summary

Researchers discovered constitutive mutations in the Escherichia coli AraC protein

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The arabinose operon regulatory protein, AraC, controls gene expression in Escherichia coli.
  • The N-terminal arm of the AraC protein is crucial for its regulatory function but is disordered in the absence of arabinose.
  • Previous studies characterized the AraC dimerization domain structure, particularly in the presence of arabinose.

Purpose of the Study:

  • To identify and characterize constitutive mutations in the N-terminal arm of the AraC protein.
  • To elucidate the molecular mechanism by which a specific mutation (N16D) leads to constitutive activity.
  • To investigate the role of charge-charge interactions in AraC protein regulation.

Main Methods:

  • Site-directed mutagenesis was used to introduce mutations into the N-terminal arm of the AraC protein.

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  • The constitutive activity of mutated AraC proteins was assessed by monitoring transcription activation.
  • Structural and interaction analyses were proposed to explain the observed mutational effects.
  • Main Results:

    • A novel constitutive mutation, N16D, was identified in the N-terminal arm of the AraC protein.
    • The N16D mutation introduces a negative charge at residue 16, potentially forming a charge-charge interaction network with Lys-43 and Arg-99.
    • Mutations of Lys-43 and Arg-99 to alanine reduced or abolished the constitutive activity conferred by the N16D mutation.

    Conclusions:

    • The N16D mutation stabilizes the N-terminal arm in a conformation that activates transcription even without arabinose.
    • A charge-charge interaction network involving Asp-16, Lys-43, and Arg-99 is proposed to mediate this constitutive activation.
    • These findings provide insights into the allosteric regulation of the AraC protein and transcriptional activation.