Chaperonin cpn60 from Escherichia coli protects the mitochondrial enzyme rhodanese against heat inactivation and supports folding at elevated temperatures.

The chaperonin protein cpn60 from Escherichia coli protects the monomeric, mitochondrial enzyme rhodanese (thiosulfate:cyanide sulfurtransferase, EC 2.8.1.1) against heat inactivation. The thermal inactivation of rhodanese was studied for four different states of the enzyme: native, refolded, bound to cpn60 in the form of a binary complex formed from unfolded rhodanese, and a thermally perturbed state. Thermal stabilization is observed in a range of temperatures from 25 to 48 degrees C. Rhodanese that had been inactivated by incubation at 48 degrees C, in the presence of cpn60 can be reactivated at 25 degrees C, upon addition of cpn10, K+, and MgATP. A recovery of about 80% was achieved after 1 h of the addition of those components. Thus, the enzyme is protected against heat inactivation and kept in a reactivable form if inactivation is attempted using the binary complex formed between rhodanese folding intermediate(s) and cpn60. The chaperonin-assisted refolding of urea-denatured rhodanese is dependent on the temperature of the refolding reaction. However, optimal chaperonin assisted refolding of rhodanese observed at 25 degrees C, which is achieved upon addition of cpn10 and ATP to the cpn60-rhodanese complex, is independent of the temperature of preincubation of the complex, that was formed previously at low temperature. The results are in agreement with a model in which the chaperonin cpn60 interacts with partly folded intermediates by forming a binary complex which is stable to elevated temperatures. In addition, it appears that native rhodanese can be thermally perturbed to produce a state different from that achieved by denaturation that can interact with cpn60.

• Publication year

  1992

• Venue

  Journal of Biological Chemistry

• Publication date

  1992-09-05

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0021-9258(19)37089-9PMID 1355476
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• cpn60 forms a stable binary complex with partly folded rhodanese intermediates, and native rhodanese can be converted into a distinct thermally perturbed state that also interacts with cpn60.

  Confidence 0.91

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• Chaperonin-assisted refolding of urea-denatured rhodanese depends on the refolding temperature, with the best recovery at 25 degrees C.

  Confidence 0.93

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• Heat-inactivated rhodanese can be reactivated at 25 degrees C by adding cpn10, K+, and MgATP, with about 80% recovery after 1 hour.

  Confidence 0.96

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• cpn60 protects rhodanese against heat inactivation over the 25 to 48 degrees C range.

  Confidence 0.98

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review

• binary complex

  molecular complex

  A complex formed between cpn60 and rhodanese folding intermediates.

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• chaperonin-assisted refolding

  process

  Refolding of denatured rhodanese promoted by cpn60 together with cpn10 and ATP.

  Aliases: chaperonin assisted refolding

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• cpn10

  protein

  The co-chaperonin added to the cpn60-rhodanese complex during reactivation and refolding reactions.

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• cpn60

  protein

  An Escherichia coli chaperonin that binds rhodanese during the experiments.

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• heat inactivation

  process

  Loss of rhodanese activity caused by incubation at elevated temperature.

  Aliases: thermal inactivation

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• k+

  ion

  Potassium ion added to support the rhodanese reactivation reaction.

  Aliases: potassium ion

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• mgatp

  cofactor

  Magnesium-bound ATP used as a reaction component in the reactivation and folding assays.

  Aliases: ATP with Mg2+

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• rhodanese

  enzyme

  The mitochondrial thiosulfate:cyanide sulfurtransferase used as the substrate enzyme in the refolding and inactivation assays.

  Aliases: thiosulfate:cyanide sulfurtransferase

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review
• thermally perturbed state

  protein state

  A heat-altered form of native rhodanese that can interact with cpn60.

  Aliases: perturbed state

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewKiller Whale (322360f1c1) reviewAK (4715169a40) review

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