title: Chaperone-like activity of Mycobacterium tuberculosis Hsp16.3 does not require its intact (native) structures.
abstract: Small heat shock proteins (sHsps) were found to exhibit efficient chaperone-like activities under stress conditions although their native structures are severely disturbed. Here, using an alternative approach (site-directed mutagenesis), we obtained two structurally and functionally distinct Mycobacterium tuberculosis Hsp16.3 single-site mutant proteins. The G59W mutant protein (with Gly59 substituted by Trp) is capable of exhibiting efficient chaperone-like activity even under non-stress conditions although its secondary, tertiary, and quaternary structures are very different from that of the wild type protein. By contrast, the G59A mutant protein (with Gly59 substituted by Ala) resembles with the wild type protein in structure and function. These observations suggest that the Gly59 of the Hsp16.3 protein is critical for its folding and assembly. In particular, we propose that the exhibition of chaperone-like activity for Hsp16.3 does not require its intact (native) structures but requires the disturbance of its native structures (i.e., the native structure-disturbed Hsp16.3 retains its chaperone-like activity or even becomes more active). In addition, the behavior of such an active mutant protein (G59W) also strongly supports our previous suggestion that Hsp16.3 exhibits chaperone-like activity via oligomeric dissociation.
authors.affiliation: Beijing Tuberculosis and Thoracic Tumor Institute, Beijing, 101149, China.
authors.forename: Xiaoyou
authors.initials: X
authors.lastname: Chen
authors.forename: Xinmiao
authors.initials: X
authors.lastname: Fu
authors.forename: Yu
authors.initials: Y
authors.lastname: Ma
authors.forename: Zengyi
authors.initials: Z
authors.lastname: Chang
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018833:Chaperonins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D014357:Trypsin / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C431133:Hsp16.3 protein, Mycobacterium tuberculosis; D018832:Molecular Chaperones; D014357:Trypsin; D018833:Chaperonins