The Functional Differences between the GroEL Chaperonin of <i>Escherichia coli</i> and the HtpB Chaperonin of <i>Legionella pneumophila</i> Can Be Mapped to Specific Amino Acid Residues
Group I chaperonins are a highly conserved family of essential proteins that self-assemble into molecular nanoboxes that mediate the folding of cytoplasmic proteins in bacteria and organelles. GroEL, the chaperonin of <i>Escherichia coli</i>, is the archetype of the family. Protein foldi...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-12-01
|
| Series: | Biomolecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-273X/12/1/59 |
| _version_ | 1797495463617757184 |
|---|---|
| author | Karla N. Valenzuela-Valderas Gabriel Moreno-Hagelsieb John R. Rohde Rafael A. Garduño |
| author_facet | Karla N. Valenzuela-Valderas Gabriel Moreno-Hagelsieb John R. Rohde Rafael A. Garduño |
| author_sort | Karla N. Valenzuela-Valderas |
| collection | DOAJ |
| description | Group I chaperonins are a highly conserved family of essential proteins that self-assemble into molecular nanoboxes that mediate the folding of cytoplasmic proteins in bacteria and organelles. GroEL, the chaperonin of <i>Escherichia coli</i>, is the archetype of the family. Protein folding-independent functions have been described for numerous chaperonins, including HtpB, the chaperonin of the bacterial pathogen <i>Legionella pneumophila</i>. Several protein folding-independent functions attributed to HtpB are not shared by GroEL, suggesting that differences in the amino acid (aa) sequence between these two proteins could correlate with functional differences. GroEL and HtpB differ in 137 scattered aa positions. Using the Evolutionary Trace (ET) bioinformatics method, site-directed mutagenesis, and a functional reporter test based upon a yeast-two-hybrid interaction with the eukaryotic protein ECM29, it was determined that out of those 137 aa, ten (M68, M212, S236, K298, N507 and the cluster AEHKD in positions 471-475) were involved in the interaction of HtpB with ECM29. GroEL was completely unable to interact with ECM29, but when GroEL was modified at those 10 aa positions, to display the HtpB aa, it acquired a weak ability to interact with ECM29. This constitutes proof of concept that the unique functional abilities of HtpB can be mapped to specific aa positions. |
| first_indexed | 2024-03-10T01:50:00Z |
| format | Article |
| id | doaj.art-5f4b02a398a541d98eb6c17be5591264 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T01:50:00Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-5f4b02a398a541d98eb6c17be55912642023-11-23T13:06:26ZengMDPI AGBiomolecules2218-273X2021-12-011215910.3390/biom12010059The Functional Differences between the GroEL Chaperonin of <i>Escherichia coli</i> and the HtpB Chaperonin of <i>Legionella pneumophila</i> Can Be Mapped to Specific Amino Acid ResiduesKarla N. Valenzuela-Valderas0Gabriel Moreno-Hagelsieb1John R. Rohde2Rafael A. Garduño3Department of Microbiology and Immunology, Dalhousie University, Sir Charles Tupper Medical Building, 7th Floor, 5850 College Street, Halifax, NS B3H 1X5, CanadaDepartment of Biology, Room BA441, Wilfrid Laurier University, 75 University Avenue West, Waterloo, ON N2L 3C5, CanadaDepartment of Microbiology and Immunology, Dalhousie University, Sir Charles Tupper Medical Building, 7th Floor, 5850 College Street, Halifax, NS B3H 1X5, CanadaDepartment of Microbiology and Immunology, Dalhousie University, Sir Charles Tupper Medical Building, 7th Floor, 5850 College Street, Halifax, NS B3H 1X5, CanadaGroup I chaperonins are a highly conserved family of essential proteins that self-assemble into molecular nanoboxes that mediate the folding of cytoplasmic proteins in bacteria and organelles. GroEL, the chaperonin of <i>Escherichia coli</i>, is the archetype of the family. Protein folding-independent functions have been described for numerous chaperonins, including HtpB, the chaperonin of the bacterial pathogen <i>Legionella pneumophila</i>. Several protein folding-independent functions attributed to HtpB are not shared by GroEL, suggesting that differences in the amino acid (aa) sequence between these two proteins could correlate with functional differences. GroEL and HtpB differ in 137 scattered aa positions. Using the Evolutionary Trace (ET) bioinformatics method, site-directed mutagenesis, and a functional reporter test based upon a yeast-two-hybrid interaction with the eukaryotic protein ECM29, it was determined that out of those 137 aa, ten (M68, M212, S236, K298, N507 and the cluster AEHKD in positions 471-475) were involved in the interaction of HtpB with ECM29. GroEL was completely unable to interact with ECM29, but when GroEL was modified at those 10 aa positions, to display the HtpB aa, it acquired a weak ability to interact with ECM29. This constitutes proof of concept that the unique functional abilities of HtpB can be mapped to specific aa positions.https://www.mdpi.com/2218-273X/12/1/59chaperoninGroELevolutionary traceECM29yeast-two-hybridLegionella |
| spellingShingle | Karla N. Valenzuela-Valderas Gabriel Moreno-Hagelsieb John R. Rohde Rafael A. Garduño The Functional Differences between the GroEL Chaperonin of <i>Escherichia coli</i> and the HtpB Chaperonin of <i>Legionella pneumophila</i> Can Be Mapped to Specific Amino Acid Residues Biomolecules chaperonin GroEL evolutionary trace ECM29 yeast-two-hybrid Legionella |
| title | The Functional Differences between the GroEL Chaperonin of <i>Escherichia coli</i> and the HtpB Chaperonin of <i>Legionella pneumophila</i> Can Be Mapped to Specific Amino Acid Residues |
| title_full | The Functional Differences between the GroEL Chaperonin of <i>Escherichia coli</i> and the HtpB Chaperonin of <i>Legionella pneumophila</i> Can Be Mapped to Specific Amino Acid Residues |
| title_fullStr | The Functional Differences between the GroEL Chaperonin of <i>Escherichia coli</i> and the HtpB Chaperonin of <i>Legionella pneumophila</i> Can Be Mapped to Specific Amino Acid Residues |
| title_full_unstemmed | The Functional Differences between the GroEL Chaperonin of <i>Escherichia coli</i> and the HtpB Chaperonin of <i>Legionella pneumophila</i> Can Be Mapped to Specific Amino Acid Residues |
| title_short | The Functional Differences between the GroEL Chaperonin of <i>Escherichia coli</i> and the HtpB Chaperonin of <i>Legionella pneumophila</i> Can Be Mapped to Specific Amino Acid Residues |
| title_sort | functional differences between the groel chaperonin of i escherichia coli i and the htpb chaperonin of i legionella pneumophila i can be mapped to specific amino acid residues |
| topic | chaperonin GroEL evolutionary trace ECM29 yeast-two-hybrid Legionella |
| url | https://www.mdpi.com/2218-273X/12/1/59 |
| work_keys_str_mv | AT karlanvalenzuelavalderas thefunctionaldifferencesbetweenthegroelchaperoninofiescherichiacoliiandthehtpbchaperoninofilegionellapneumophilaicanbemappedtospecificaminoacidresidues AT gabrielmorenohagelsieb thefunctionaldifferencesbetweenthegroelchaperoninofiescherichiacoliiandthehtpbchaperoninofilegionellapneumophilaicanbemappedtospecificaminoacidresidues AT johnrrohde thefunctionaldifferencesbetweenthegroelchaperoninofiescherichiacoliiandthehtpbchaperoninofilegionellapneumophilaicanbemappedtospecificaminoacidresidues AT rafaelagarduno thefunctionaldifferencesbetweenthegroelchaperoninofiescherichiacoliiandthehtpbchaperoninofilegionellapneumophilaicanbemappedtospecificaminoacidresidues AT karlanvalenzuelavalderas functionaldifferencesbetweenthegroelchaperoninofiescherichiacoliiandthehtpbchaperoninofilegionellapneumophilaicanbemappedtospecificaminoacidresidues AT gabrielmorenohagelsieb functionaldifferencesbetweenthegroelchaperoninofiescherichiacoliiandthehtpbchaperoninofilegionellapneumophilaicanbemappedtospecificaminoacidresidues AT johnrrohde functionaldifferencesbetweenthegroelchaperoninofiescherichiacoliiandthehtpbchaperoninofilegionellapneumophilaicanbemappedtospecificaminoacidresidues AT rafaelagarduno functionaldifferencesbetweenthegroelchaperoninofiescherichiacoliiandthehtpbchaperoninofilegionellapneumophilaicanbemappedtospecificaminoacidresidues |