Related Experiment Video

Updated: Aug 26, 2026

Escherichia coli -Based Complementation Assay to Study the Chaperone Function of Heat Shock Protein 70
07:14

Escherichia coli -Based Complementation Assay to Study the Chaperone Function of Heat Shock Protein 70

Published on: March 8, 2024

73-kDa molecular chaperone HSP73 is a direct target of antibiotic gentamicin

Toshio Miyazaki1, Ryo Sagawa, Takenori Honma

  • 1Department of Material-process Engineering and Applied Chemistry for Environment, Akita University Faculty of Engineering and Resource Science, 1-1 Tegata Gakuen Town, Akita City 010-8502, Japan.

The Journal of Biological Chemistry
|February 18, 2004
PubMed

Insights

Gentamicin (GM) antibiotic binds to the 73-kDa heat shock protein (HSP73), reducing its chaperone activity. This interaction may contribute to gentamicin-induced kidney toxicity.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Nephrology

Background:

  • Gentamicin (GM) is a widely used antibiotic.
  • The specific protein targets of GM remain incompletely understood.
  • Understanding GM's molecular interactions is crucial for elucidating its toxicity mechanisms.

Purpose of the Study:

  • To identify the specific binding protein of gentamicin.
  • To investigate the functional consequences of gentamicin binding to its target protein.
  • To explore the role of this interaction in gentamicin-induced kidney injury.

Main Methods:

  • Gentamicin-affinity chromatography was used to isolate binding proteins from porcine kidney cytosol.
  • Immunoblotting and CD spectroscopy were employed to identify and characterize the binding protein.
  • In vitro assays assessed the effect of gentamicin on HSP73 chaperone activity.
  • Limited proteolysis mapped the gentamicin binding site on HSP73.
  • Immunohistochemistry examined the co-localization of HSP73 and gentamicin in rat kidneys.

Main Results:

  • A 73-kDa protein, identified as heat shock protein 73 (HSP73), specifically bound to gentamicin.
  • Gentamicin binding induced a conformational change in HSP73.
  • Gentamicin suppressed the in vitro chaperone activity of HSP73.
  • The gentamicin binding site on HSP73 was localized to the C-terminal peptide-binding domain.
  • HSP73 and gentamicin were co-localized in lysosomes of rat kidneys exhibiting gentamicin-induced acute tubular injury.

Conclusions:

  • Gentamicin specifically binds to HSP73, reducing its chaperone activity.
  • This interaction may play a role in the pathogenesis of gentamicin-induced nephrotoxicity.
  • Targeting the HSP73-gentamicin interaction could offer novel therapeutic strategies for mitigating gentamicin toxicity.

More Related Videos

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay06:51

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay

Published on: July 21, 2021

Intracellular Refolding Assay07:18

Intracellular Refolding Assay

Published on: January 24, 2012

Related Concept Videos

Inhibitors of Bacterial Protein Synthesis01:25

Inhibitors of Bacterial Protein Synthesis

Aminoglycosides constitute a highly potent class of bactericidal antibiotics that exert their antimicrobial effects by targeting the bacterial ribosome, specifically disrupting protein synthesis. These polycationic molecules consist of amino-modified sugars linked via glycosidic bonds to an aminocyclitol core such as 2-deoxystreptamine or streptamine. Their strong positive charges facilitate tight binding to the negatively charged phosphate backbone of ribosomal RNA (rRNA), primarily at the 16S...
Bacterial Protein Maturation01:26

Bacterial Protein Maturation

Bacterial protein maturation is a tightly regulated process that ensures newly synthesized polypeptides achieve correct functional conformations. This maturation involves a series of modifications, folding events, and quality control steps, often assisted by specialized chaperone proteins.N-Terminal ModificationsThe maturation of bacterial polypeptides begins cotranslationally as the polypeptide exits the ribosome. The first amino acid, N-formylmethionine (fMet), is typically modified at the...
Molecular Chaperones and Protein Folding03:00

Molecular Chaperones and Protein Folding

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.
The...
Determination of Multiple Dosing Parameters: Steady-State, Minimum and Maximum Concentrations01:15

Determination of Multiple Dosing Parameters: Steady-State, Minimum and Maximum Concentrations

Gentamicin, an aminoglycoside antibiotic, is commonly administered via intermittent intravenous infusion to treat severe infections. An intermittent one-hour infusion of gentamicin, administered at eight-hour intervals, allows for precise control of plasma drug concentrations, minimizing toxicity while ensuring therapeutic efficacy. Pharmacokinetic principles govern the dynamics of plasma concentrations and can be mathematically described using specific equations.The plasma drug concentration...
Inhibitors of Gram-positive Cell Wall Synthesis01:23

Inhibitors of Gram-positive Cell Wall Synthesis

Bacterial cell walls are typically rigid structures composed mainly of peptidoglycan, a mesh-like polymer that provides mechanical strength and maintains cell shape. The synthesis of peptidoglycan is a crucial process in bacterial growth and serves as a primary target for many antibiotics.Mechanism of Action of Beta-Lactam AntibioticsBeta-lactam antibiotics, such as penicillin, inhibit peptidoglycan synthesis in actively growing cells. These antibiotics share a characteristic four-membered...
Mechanism of Antibiotic Resistance in MRSA01:25

Mechanism of Antibiotic Resistance in MRSA

Antibiotic resistance in bacteria arises when microorganisms evolve the ability to withstand drugs designed to kill them or inhibit their growth, rendering once-effective treatments useless. This phenomenon, driven by genetic change and selection under antibiotic exposure, poses a profound threat to modern medicine. Mechanisms include drug-inactivating enzymes (e.g., β-lactamases), efflux pumps that eject antibiotics, mutations altering antibiotic targets, decreased drug uptake, and acquisition...