A Dual Role for the N-terminal Region of Mycobacterium tuberculosis Hsp16.3 in Self-oligomerization and Binding Denaturing Substrate Proteins
| Autor: | Abuduaini Abulimiti, Yang Song, Chong Liu, Yang Cao, Wangwang Jiao, Xuefeng Zhang, Xinmiao Fu, Hui Zhang, Zengyi Chang |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Protein Denaturation
Methanococcus Chaperonins Stereochemistry Dimer Protein subunit Plasma protein binding Biology Biochemistry Oligomer chemistry.chemical_compound Bacterial Proteins Mutant protein Amino Acid Sequence Binding site Molecular Biology Sequence Deletion Binding Sites Mycobacterium tuberculosis Cell Biology biology.organism_classification Fusion protein Protein Subunits chemistry Mutagenesis Site-Directed Dimerization Molecular Chaperones Protein Binding |
| Zdroj: | Journal of Biological Chemistry. 280:6337-6348 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m406319200 |
| Popis: | The N-terminal regions, which are highly variable in small heat-shock proteins, were found to be structurally disordered in all the 24 subunits of Methanococcus jannaschii Hsp16.5 oligomer and half of the 12 subunits of wheat Hsp16.9 oligomer. The structural and functional roles of the corresponding region (potentially disordered) in Mycobacterium tuberculosis Hsp16.3, existing as nonamers, were investigated in this work. The data demonstrate that the mutant Hsp16.3 protein with 35 N-terminal residues removed (DeltaN35) existed as trimers/dimers rather than as nonamers, failing to bind the hydrophobic probe (1,1'-bi(4-anilino)naphthalene-5,5'-disulfonic acid) and exhibiting no chaperone-like activity. Nevertheless, another mutant protein with the C-terminal extension (of nine residues) removed, although existing predominantly as dimers, exhibited efficient chaperone-like activity even at room temperatures, indicating that pre-existence as nonamers is not a prerequisite for its chaperone-like activity. Meanwhile, the mutant protein with both the N- and C-terminal ends removed fully exists as a dimer lacking any chaperone-like activity. Furthermore, the N-terminal region alone, either as a synthesized peptide or in fusion protein with glutathione S-transferase, was capable of interacting with denaturing proteins. These observations strongly suggest that the N-terminal region of Hsp16.3 is not only involved in self-oligomerization but also contains the critical site for substrate binding. Such a dual role for the N-terminal region would provide an effective mechanism for the small heat-shock protein to modulate its chaperone-like activity through oligomeric dissociation/reassociation. In addition, this study demonstrated that the wild-type protein was able to form heterononamers with DeltaN35 via subunit exchange at a subunit ratio of 2:1. This implies that the 35 N-terminal residues in three of the nine subunits in the wild-type nonamer are not needed for the assembly of nonamers from trimers and are thus probably structurally disordered. |
| Databáze: | OpenAIRE |
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