Isolation of genes encoding heat shock protein 70 (hsp70s) from the coelacanth, Latimeria chalumnae
- Modisakeng, Keoagile W, Dorrington, Rosemary A, Blatch, Gregory L
- Authors: Modisakeng, Keoagile W , Dorrington, Rosemary A , Blatch, Gregory L
- Date: 2004
- Language: English
- Type: Article
- Identifier: vital:6459 , http://hdl.handle.net/10962/d1005788
- Description: Under stress conditions, proteins unfold or misfold, leading to the formation of aggregates. Molecular chaperones can be defined as proteins that facilitate the correct folding of other proteins, so that they attain a stable tertiary structure. In addition, they promote the refolding and degradation of denatured proteins after cellular stress. Heat shock proteins form one of the main classes of molecular chaperones. We are interested in determining if the genome of the coelacanth (Latimeria chalumnae) encodes a heat shock protein-based cytoprotection mechanism. We have isolated 50 kb and larger coelacanth genomic DNA from frozen skin tissue of L. chalumnae. From the alignments of several fish Hsp70 proteins, conserved regions at the N- and C-termini were identified. Codon usage tables were constructed from published coelacanth genes and degenerate primers were designed to isolate the full-length hsp70 gene and regions encoding the ATPase and the peptide binding domains. Since it is known that the tilapia and Fugu inducible hsp70 genes are intronless, we proceeded on the assumption that a coelacanth inducible hsp70 would also be intronless. A large fragment (1840 bp) encoding most of a coelacanth Hsp70 protein, and two partial fragments encoding a coelacanth Hsp70ATPase domain (1048 bp) and peptide binding domain (873 bp), were isolated by polymerase chain reaction amplification. Protein sequences translated from all the nucleotide sequences were closely identical to typical Hsp70s. This is the first study to provide evidence for a cytoprotection mechanism in the coelacanth involving an inducible Hsp70.
- Full Text:
- Date Issued: 2004
- Authors: Modisakeng, Keoagile W , Dorrington, Rosemary A , Blatch, Gregory L
- Date: 2004
- Language: English
- Type: Article
- Identifier: vital:6459 , http://hdl.handle.net/10962/d1005788
- Description: Under stress conditions, proteins unfold or misfold, leading to the formation of aggregates. Molecular chaperones can be defined as proteins that facilitate the correct folding of other proteins, so that they attain a stable tertiary structure. In addition, they promote the refolding and degradation of denatured proteins after cellular stress. Heat shock proteins form one of the main classes of molecular chaperones. We are interested in determining if the genome of the coelacanth (Latimeria chalumnae) encodes a heat shock protein-based cytoprotection mechanism. We have isolated 50 kb and larger coelacanth genomic DNA from frozen skin tissue of L. chalumnae. From the alignments of several fish Hsp70 proteins, conserved regions at the N- and C-termini were identified. Codon usage tables were constructed from published coelacanth genes and degenerate primers were designed to isolate the full-length hsp70 gene and regions encoding the ATPase and the peptide binding domains. Since it is known that the tilapia and Fugu inducible hsp70 genes are intronless, we proceeded on the assumption that a coelacanth inducible hsp70 would also be intronless. A large fragment (1840 bp) encoding most of a coelacanth Hsp70 protein, and two partial fragments encoding a coelacanth Hsp70ATPase domain (1048 bp) and peptide binding domain (873 bp), were isolated by polymerase chain reaction amplification. Protein sequences translated from all the nucleotide sequences were closely identical to typical Hsp70s. This is the first study to provide evidence for a cytoprotection mechanism in the coelacanth involving an inducible Hsp70.
- Full Text:
- Date Issued: 2004
Molecular chaperones in biology, medicine and protein biotechnology
- Boshoff, Aileen, Nicoll, William S, Hennessy, Fritha, Ludewig, M H, Daniel, Sheril, Modisakeng, Keoagile W, Shonhai, Addmore, McNamara, Caryn, Bradley, Graeme, Blatch, Gregory L
- Authors: Boshoff, Aileen , Nicoll, William S , Hennessy, Fritha , Ludewig, M H , Daniel, Sheril , Modisakeng, Keoagile W , Shonhai, Addmore , McNamara, Caryn , Bradley, Graeme , Blatch, Gregory L
- Date: 2004
- Language: English
- Type: Article
- Identifier: vital:6457 , http://hdl.handle.net/10962/d1004479
- Description: Molecular chaperones consist of several highly conserved families of proteins, many of which consist of heat shock proteins. The primary function of molecular chaperones is to facilitate the folding or refolding of proteins, and therefore they play an important role in diverse cellular processes including protein synthesis, protein translocation, and the refolding or degradation of proteins after cell stress. Cells are often exposed to different stressors, resulting in protein misfolding and aggregation. It is now well established that the levels of certain molecular chaperones are elevated during stress to provide protection to the cell. The focus of this review is on the impact of molecular chaperones in biology, medicine and protein biotechnology, and thus covers both fundamental and applied aspects of chaperone biology. Attention is paid to the functions and applications of molecular chaperones from bacterial and eukaryotic cells, focusing on the heat shock proteins 90 (Hsp90), 70 (Hsp70) and 40 (Hsp40) classes of chaperones, respectively. The role of these classes of chaperones in human diseases is discussed, as well as the parts played by chaperones produced by the causative agents of malaria and trypanosomiasis. Recent advances have seen the application of chaperones in improving the yields of a particular target protein in recombinant protein production. The prospects for the targeted use of molecular chaperones for the over-production of recombinant proteins is critically reviewed, and current research on these chaperones at Rhodes University is also discussed.
- Full Text:
- Date Issued: 2004
- Authors: Boshoff, Aileen , Nicoll, William S , Hennessy, Fritha , Ludewig, M H , Daniel, Sheril , Modisakeng, Keoagile W , Shonhai, Addmore , McNamara, Caryn , Bradley, Graeme , Blatch, Gregory L
- Date: 2004
- Language: English
- Type: Article
- Identifier: vital:6457 , http://hdl.handle.net/10962/d1004479
- Description: Molecular chaperones consist of several highly conserved families of proteins, many of which consist of heat shock proteins. The primary function of molecular chaperones is to facilitate the folding or refolding of proteins, and therefore they play an important role in diverse cellular processes including protein synthesis, protein translocation, and the refolding or degradation of proteins after cell stress. Cells are often exposed to different stressors, resulting in protein misfolding and aggregation. It is now well established that the levels of certain molecular chaperones are elevated during stress to provide protection to the cell. The focus of this review is on the impact of molecular chaperones in biology, medicine and protein biotechnology, and thus covers both fundamental and applied aspects of chaperone biology. Attention is paid to the functions and applications of molecular chaperones from bacterial and eukaryotic cells, focusing on the heat shock proteins 90 (Hsp90), 70 (Hsp70) and 40 (Hsp40) classes of chaperones, respectively. The role of these classes of chaperones in human diseases is discussed, as well as the parts played by chaperones produced by the causative agents of malaria and trypanosomiasis. Recent advances have seen the application of chaperones in improving the yields of a particular target protein in recombinant protein production. The prospects for the targeted use of molecular chaperones for the over-production of recombinant proteins is critically reviewed, and current research on these chaperones at Rhodes University is also discussed.
- Full Text:
- Date Issued: 2004
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