The amino‐terminal domain of Mycobacterium tuberculosis ClpB protein plays a crucial role in its substrate disaggregation activity
Mycobacterium tuberculosis (Mtb) is known to persist in extremely hostile environments within host macrophages. The ability to withstand such proteotoxic stress comes from its highly conserved molecular chaperone machinery. ClpB, a unique member of the AAA+ family of chaperones, is responsible for r...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2018-10-01
|
| Series: | FEBS Open Bio |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/2211-5463.12509 |
| _version_ | 1798014982997147648 |
|---|---|
| author | Prajna Tripathi Priyanka Parijat Virendra Kumar Patel Janendra K. Batra |
| author_facet | Prajna Tripathi Priyanka Parijat Virendra Kumar Patel Janendra K. Batra |
| author_sort | Prajna Tripathi |
| collection | DOAJ |
| description | Mycobacterium tuberculosis (Mtb) is known to persist in extremely hostile environments within host macrophages. The ability to withstand such proteotoxic stress comes from its highly conserved molecular chaperone machinery. ClpB, a unique member of the AAA+ family of chaperones, is responsible for resolving aggregates in Mtb and many other bacterial pathogens. Mtb produces two isoforms of ClpB, a full length and an N‐terminally truncated form (ClpB∆N), with the latter arising from an internal translation initiation site. It is not clear why this internal start site is conserved and what role the N‐terminal domain (NTD) of Mtb ClpB plays in its function. In the current study, we functionally characterized and compared the two isoforms of Mtb ClpB. We found the NTD to be dispensable for oligomerization, ATPase activity and prevention of aggregation activity of ClpB. Both ClpB and ClpB∆N were found to be capable of resolubilizing protein aggregates. However, the efficiency of ClpB∆N at resolubilizing higher order aggregates was significantly lower than that of ClpB. Further, ClpB∆N exhibited reduced affinity for substrates as compared to ClpB. We also demonstrated that the surface of the NTD of Mtb ClpB has a hydrophobic groove that contains four hydrophobic residues: L97, L101, F140 and V141. These residues act as initial contacts for the substrate and are crucial for stable interaction between ClpB and highly aggregated substrates. |
| first_indexed | 2024-04-11T15:26:59Z |
| format | Article |
| id | doaj.art-915bb9c4739a4a288ef92b69bc81e01e |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-11T15:26:59Z |
| publishDate | 2018-10-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-915bb9c4739a4a288ef92b69bc81e01e2022-12-22T04:16:14ZengWileyFEBS Open Bio2211-54632018-10-018101669169010.1002/2211-5463.12509The amino‐terminal domain of Mycobacterium tuberculosis ClpB protein plays a crucial role in its substrate disaggregation activityPrajna Tripathi0Priyanka Parijat1Virendra Kumar Patel2Janendra K. Batra3National Institute of Immunology New Delhi IndiaNational Institute of Immunology New Delhi IndiaNational Institute of Immunology New Delhi IndiaNational Institute of Immunology New Delhi IndiaMycobacterium tuberculosis (Mtb) is known to persist in extremely hostile environments within host macrophages. The ability to withstand such proteotoxic stress comes from its highly conserved molecular chaperone machinery. ClpB, a unique member of the AAA+ family of chaperones, is responsible for resolving aggregates in Mtb and many other bacterial pathogens. Mtb produces two isoforms of ClpB, a full length and an N‐terminally truncated form (ClpB∆N), with the latter arising from an internal translation initiation site. It is not clear why this internal start site is conserved and what role the N‐terminal domain (NTD) of Mtb ClpB plays in its function. In the current study, we functionally characterized and compared the two isoforms of Mtb ClpB. We found the NTD to be dispensable for oligomerization, ATPase activity and prevention of aggregation activity of ClpB. Both ClpB and ClpB∆N were found to be capable of resolubilizing protein aggregates. However, the efficiency of ClpB∆N at resolubilizing higher order aggregates was significantly lower than that of ClpB. Further, ClpB∆N exhibited reduced affinity for substrates as compared to ClpB. We also demonstrated that the surface of the NTD of Mtb ClpB has a hydrophobic groove that contains four hydrophobic residues: L97, L101, F140 and V141. These residues act as initial contacts for the substrate and are crucial for stable interaction between ClpB and highly aggregated substrates.https://doi.org/10.1002/2211-5463.12509Clp proteinsdisaggregationDnaKheat shock proteins (HSP)M. tuberculosisRv0384c |
| spellingShingle | Prajna Tripathi Priyanka Parijat Virendra Kumar Patel Janendra K. Batra The amino‐terminal domain of Mycobacterium tuberculosis ClpB protein plays a crucial role in its substrate disaggregation activity FEBS Open Bio Clp proteins disaggregation DnaK heat shock proteins (HSP) M. tuberculosis Rv0384c |
| title | The amino‐terminal domain of Mycobacterium tuberculosis ClpB protein plays a crucial role in its substrate disaggregation activity |
| title_full | The amino‐terminal domain of Mycobacterium tuberculosis ClpB protein plays a crucial role in its substrate disaggregation activity |
| title_fullStr | The amino‐terminal domain of Mycobacterium tuberculosis ClpB protein plays a crucial role in its substrate disaggregation activity |
| title_full_unstemmed | The amino‐terminal domain of Mycobacterium tuberculosis ClpB protein plays a crucial role in its substrate disaggregation activity |
| title_short | The amino‐terminal domain of Mycobacterium tuberculosis ClpB protein plays a crucial role in its substrate disaggregation activity |
| title_sort | amino terminal domain of mycobacterium tuberculosis clpb protein plays a crucial role in its substrate disaggregation activity |
| topic | Clp proteins disaggregation DnaK heat shock proteins (HSP) M. tuberculosis Rv0384c |
| url | https://doi.org/10.1002/2211-5463.12509 |
| work_keys_str_mv | AT prajnatripathi theaminoterminaldomainofmycobacteriumtuberculosisclpbproteinplaysacrucialroleinitssubstratedisaggregationactivity AT priyankaparijat theaminoterminaldomainofmycobacteriumtuberculosisclpbproteinplaysacrucialroleinitssubstratedisaggregationactivity AT virendrakumarpatel theaminoterminaldomainofmycobacteriumtuberculosisclpbproteinplaysacrucialroleinitssubstratedisaggregationactivity AT janendrakbatra theaminoterminaldomainofmycobacteriumtuberculosisclpbproteinplaysacrucialroleinitssubstratedisaggregationactivity AT prajnatripathi aminoterminaldomainofmycobacteriumtuberculosisclpbproteinplaysacrucialroleinitssubstratedisaggregationactivity AT priyankaparijat aminoterminaldomainofmycobacteriumtuberculosisclpbproteinplaysacrucialroleinitssubstratedisaggregationactivity AT virendrakumarpatel aminoterminaldomainofmycobacteriumtuberculosisclpbproteinplaysacrucialroleinitssubstratedisaggregationactivity AT janendrakbatra aminoterminaldomainofmycobacteriumtuberculosisclpbproteinplaysacrucialroleinitssubstratedisaggregationactivity |