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Related Concept Videos

Molecular Chaperones and Protein Folding03:00

Molecular Chaperones and Protein Folding

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The native conformation of a protein is formed by interactions between the side chains of its constituent amino acids. When the amino acids cannot form these interactions, the protein cannot fold by itself and needs chaperones. Notably, chaperones do not relay any additional information required for the folding of polypeptides; the native conformation of a protein is determined solely by its amino acid sequence. Chaperones catalyze protein folding without being a part of the folded protein.
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Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
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Protein Kinases and Phosphatases02:54

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Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
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Phosphorylation01:02

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The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
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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.
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Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...
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Related Experiment Video

Updated: May 24, 2025

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
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Phosphorylation-State Modulated Binding of HSP70: Structural Insights and Compensatory Protein Engineering.

Mariah Stewart1,2, Chathura Paththamperuma3, Colleen McCann1

  • 1The McAllister Heart Institute, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Biorxiv : the Preprint Server for Biology
|March 3, 2025
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Summary

Phosphorylation of heat shock protein 70 (HSP70) at T636 weakens its interaction with the E3 ligase CHIP, impacting protein quality control. This discovery offers insights into diseases linked to proteostasis imbalance.

Keywords:
CHIP (C-terminus of HSC70 interacting protein)ChaperoneCo-chaperoneHSP70PhosphorylationPost-translational modificationsProtein quality control

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Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Biochemistry

Background:

  • Protein quality control is vital for cellular homeostasis, relying on systems like the heat shock response, ubiquitin-proteasome system, and autophagy-lysosome pathway.
  • Heat shock protein 70 (HSP70) and heat shock cognate 70 (HSC70) are key chaperones managing protein folding and degradation.
  • The co-chaperone CHIP acts as an E3 ligase, targeting proteins for degradation via ubiquitination.

Purpose of the Study:

  • To investigate the effect of C-terminal phosphorylation of HSP70 on its interaction with the co-chaperone CHIP.
  • To elucidate the structural and functional consequences of HSP70 phosphorylation on chaperone-mediated protein degradation.
  • To explore the implications of these interactions for cellular homeostasis and disease pathogenesis.

Main Methods:

  • Utilized cell-free and cell-based assays, including X-ray crystallography and biolayer interferometry.
  • Employed live cell biocomplementation assays to study protein interactions in a cellular context.
  • Engineered a CHIP variant (CHIP-G132N) to assess its binding affinity to phosphorylated HSP70.

Main Results:

  • HSP70 T636 phosphorylation significantly reduces binding affinity to CHIP, favoring other co-chaperones like HOP.
  • Structural analysis revealed that phosphorylation disrupts critical hydrogen bonds, altering binding dynamics between HSP70 and CHIP.
  • While CHIP-G132N partially restored binding to phosphorylated HSP70, its efficacy was reduced in full-length constructs, suggesting complex regulatory interactions.

Conclusions:

  • Post-translational modifications, specifically phosphorylation, play a critical role in modulating chaperone-co-chaperone interactions within protein quality control networks.
  • Altered HSP70-CHIP interactions due to phosphorylation impact protein stability and degradation, with potential links to cancer and neurodegenerative diseases.
  • Understanding these molecular mechanisms provides a basis for developing novel therapeutic strategies for diseases associated with proteostasis disruption.