The Mycobacterium tuberculosis small heat shock protein Hsp16.3 exposes hydrophobic surfaces at mild conditions: conformational flexibility and molecular chaperone activity.
Hsp16.3, the alpha-crystallin-related small heat shock protein of Mycobacterium tuberculosis that is maximally expressed during the stationary phase and is a major membrane protein, has been reported to form specific trimer-of-trimers structure and to act as an effective molecular chaperone (Chang Z et al., 1996, J. Biol Chem 271:7218-7223). However, little is known about its action mechanism. In this study, Hsp16.3 conformational intermediates with dramatically increased chaperone activities were detected after treatment with very low concentrations of guanidine hydrochloride (0.05 M), urea (0.3 M), or mild heating (30 degrees C). The intermediates showed a significant increase in their capacity to bind the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS), indicating an increased exposure of hydrophobic surfaces. Interestingly, the greatest chaperone activities of Hsp16.3 were observed in the presence of 0.3 M guanidine HCl or when heated to 35 degrees C. CD spectroscopy studies revealed no significant changes in protein secondary and tertiary structures at these mild treatments. Our in vitro studies also indicate that long-time-heated Hsp16.3, heated even to temperatures as high as 85 degrees C, has almost the same, if not a slightly greater, chaperone activities as the native protein when cooled to room temperature and its secondary structures also almost recovered. Together, these results suggest that Hsp16.3 modulates its chaperone activity by exposing hydrophobic surfaces and that the protein structure is highly stable and flexible, thus highly adapted for its function.
Yang
H
H
Department of Biological Science and Biotechnology, Tsinghua University, Beijing, People's Republic of China.
Huang
S
S
Dai
H
H
Gong
Y
Y
Zheng
C
C
Chang
Z
Z
D000817:Anilino Naphthalenesulfonates; D002942:Circular Dichroism; D005456:Fluorescent Dyes; D019791:Guanidine / Q000737:chemistry; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000378:metabolism; D007328:Insulin / Q000737:chemistry; D018832:Molecular Chaperones / Q000737:chemistry* / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D006360:Heat-Shock Proteins; D007328:Insulin; D018832:Molecular Chaperones; C027132:1-anilino-8-naphthalenesulfonate; D019791:Guanidine