Related Experiment Video

Updated: Aug 8, 2026

Purification of Hsp104, a Protein Disaggregase
07:17

Purification of Hsp104, a Protein Disaggregase

Published on: September 30, 2011

Isolation and some properties of bovine brain 100 kDa heat shock protein

H Itoh1, R Kobayashi, Y Tashima

  • 1Department of Biochemistry, Akita University School of Medicine, Japan.

The International Journal of Biochemistry
|January 1, 1990
PubMed

Insights

Researchers identified a 100 kDa heat shock protein (HSP100) in bovine brains. This protein shares a common peptide fragment with HSP90, indicating a close relationship and potential functional overlap between these heat shock proteins.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Science

Background:

  • Heat shock proteins (HSPs) play crucial roles in cellular stress response and protein homeostasis.
  • HSP100 and HSP90 are important members of the HSP family, involved in various cellular processes.
  • Understanding the relationship between different HSPs can elucidate their functions and interactions.

Purpose of the Study:

  • To purify and characterize a 100 kDa protein from bovine brains.
  • To investigate the relationship between the purified 100 kDa protein and other heat shock proteins, specifically HSP100 and HSP90.
  • To determine if the 100 kDa protein is indeed HSP100 and to explore its homology with HSP90.

Main Methods:

  • Protein purification from bovine brain tissue.
  • Antibody preparation and characterization (monospecificity).
  • Immunological cross-reactivity assays (HeLa cell HSP100).
  • Physicochemical and immunochemical property analysis.
  • Partial amino acid sequencing.
  • Peptide mapping using Staphylococcus aureus V8 protease.
  • Amino acid sequence comparison.

Main Results:

  • A 100 kDa protein was successfully purified from bovine brains.
  • The generated antibody was monospecific and cross-reacted with HeLa cell HSP100, identifying the protein as HSP100.
  • Physicochemical and immunochemical analyses supported the identification of the protein as HSP100.
  • Peptide mapping revealed a common 10 kDa core peptide between HSP100 and HSP90.
  • The 10 kDa fragment of bovine HSP100 showed high homology (21/23 amino acids) to human HSP90 fragments (38-60).

Conclusions:

  • The 100 kDa protein purified from bovine brains is identified as HSP100.
  • HSP100 and HSP90 share a conserved peptide region, suggesting evolutionary relatedness and potential functional links.
  • The high sequence homology indicates a close molecular relationship between bovine HSP100 and human HSP90.

More Related Videos

Isolation of Soluble and Insoluble PrP Oligomers in the Normal Human Brain11:29

Isolation of Soluble and Insoluble PrP Oligomers in the Normal Human Brain

Published on: October 3, 2012

Reliable Isolation of Central Nervous System Microvessels Across Five Vertebrate Groups10:35

Reliable Isolation of Central Nervous System Microvessels Across Five Vertebrate Groups

Published on: January 12, 2020

Related Concept Videos

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...
Western Blotting01:15

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
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...