Preheat treatment for Mycobacterium tuberculosis Hsp16.3: correlation between a structural phase change at 60 degrees C and a dramatic increase in chaperone-like activity

Biochem Biophys Res Commun. 2001 Jun 22;284(4):942-7. doi: 10.1006/bbrc.2001.5074.


The in vitro chaperone-like activity of Mycobacterium tuberculosis small heat shock protein Hsp16.3 was found to be dramatically enhanced to the same extent after preheat treatment at or over 60 degrees C. Structural analysis using gel filtration, native pore-gradient PAGE, nondenaturing PAGE, and far-UV CD spectroscopy consistently revealed no significant difference between the native and the preheated Hsp16.3 proteins. However, near-UV CD spectroscopy clearly demonstrated that the tertiary structure of preheated Hsp16.3 is quite similar to its native conformation, with a minor but significant difference. Further analysis using differential scanning calorimetry indicated that Hsp16.3 exhibited a structural transition near 60 degrees C. All these results together indicate that Hsp16.3 suffers a phase change at approximately 60 degrees C, which seem to remove a structural energy barrier for the protein to refold to a conformational status with increased chaperone-like activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins*
  • Calorimetry, Differential Scanning
  • Chaperonins / chemistry*
  • Chaperonins / metabolism*
  • Circular Dichroism
  • Dithiothreitol / pharmacology
  • Hot Temperature
  • Insulin / chemistry*
  • Mycobacterium tuberculosis / metabolism*
  • Protein Conformation
  • Protein Denaturation
  • Spectrophotometry, Ultraviolet
  • Thermodynamics


  • Bacterial Proteins
  • Hsp16.3 protein, Mycobacterium tuberculosis
  • Insulin
  • Chaperonins
  • Dithiothreitol