The high resolution crystal structure of a native thermostable serpin reveals the complex mechanism underpinning the stressed to relaxed transition

Kate F. Fulton, Ashley M. Buckle, Lisa D. Cabrita, James A. Irving, Rebecca E. Butcher, Ian Smith, Shane Reeve, Arthur Lesk, Stephen P. Bottomley, Jamie Rossjohn, James C. Whisstock

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Serpins fold into a native metastable state and utilize a complex conformational change to inhibit target proteases. An undesirable result of this conformational flexibility is that most inhibitory serpins are heat sensitive, forming inactive polymers at elevated temperatures. However, the prokaryote serpin, thermopin, from Thermobifida fusca is able to function in a heated environment. We have determined the 1.8 Å x-ray crystal structure of thermopin in the native, inhibitory conformation. A structural comparison with the previously determined 1.5 Å structure of cleaved thermopin provides detailed insight into the complex mechanism of conformational change in serpins. Flexibility in the shutter region and electrostatic interactions at the top of the A β-sheet (the breach) involving the C-terminal tail, a unique structural feature of thermopin, are postulated to be important for controlling inhibitory activity and triggering conformational change, respectively, in the native state. Here we have discussed the structural basis of how this serpin reconciles the thermodynamic instability necessary for function with the stability required to withstand elevated temperatures.

Original languageEnglish (US)
Pages (from-to)8435-8442
Number of pages8
JournalJournal of Biological Chemistry
Volume280
Issue number9
DOIs
StatePublished - Mar 4 2005

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Serpins
Crystal structure
Temperature
Coulomb interactions
Static Electricity
Thermodynamics
Conformations
Polymers
Peptide Hydrolases
Hot Temperature
X-Rays
X rays

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Fulton, Kate F. ; Buckle, Ashley M. ; Cabrita, Lisa D. ; Irving, James A. ; Butcher, Rebecca E. ; Smith, Ian ; Reeve, Shane ; Lesk, Arthur ; Bottomley, Stephen P. ; Rossjohn, Jamie ; Whisstock, James C. / The high resolution crystal structure of a native thermostable serpin reveals the complex mechanism underpinning the stressed to relaxed transition. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 9. pp. 8435-8442.
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abstract = "Serpins fold into a native metastable state and utilize a complex conformational change to inhibit target proteases. An undesirable result of this conformational flexibility is that most inhibitory serpins are heat sensitive, forming inactive polymers at elevated temperatures. However, the prokaryote serpin, thermopin, from Thermobifida fusca is able to function in a heated environment. We have determined the 1.8 {\AA} x-ray crystal structure of thermopin in the native, inhibitory conformation. A structural comparison with the previously determined 1.5 {\AA} structure of cleaved thermopin provides detailed insight into the complex mechanism of conformational change in serpins. Flexibility in the shutter region and electrostatic interactions at the top of the A β-sheet (the breach) involving the C-terminal tail, a unique structural feature of thermopin, are postulated to be important for controlling inhibitory activity and triggering conformational change, respectively, in the native state. Here we have discussed the structural basis of how this serpin reconciles the thermodynamic instability necessary for function with the stability required to withstand elevated temperatures.",
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Fulton, KF, Buckle, AM, Cabrita, LD, Irving, JA, Butcher, RE, Smith, I, Reeve, S, Lesk, A, Bottomley, SP, Rossjohn, J & Whisstock, JC 2005, 'The high resolution crystal structure of a native thermostable serpin reveals the complex mechanism underpinning the stressed to relaxed transition', Journal of Biological Chemistry, vol. 280, no. 9, pp. 8435-8442. https://doi.org/10.1074/jbc.M410206200

The high resolution crystal structure of a native thermostable serpin reveals the complex mechanism underpinning the stressed to relaxed transition. / Fulton, Kate F.; Buckle, Ashley M.; Cabrita, Lisa D.; Irving, James A.; Butcher, Rebecca E.; Smith, Ian; Reeve, Shane; Lesk, Arthur; Bottomley, Stephen P.; Rossjohn, Jamie; Whisstock, James C.

In: Journal of Biological Chemistry, Vol. 280, No. 9, 04.03.2005, p. 8435-8442.

Research output: Contribution to journalArticle

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AU - Fulton, Kate F.

AU - Buckle, Ashley M.

AU - Cabrita, Lisa D.

AU - Irving, James A.

AU - Butcher, Rebecca E.

AU - Smith, Ian

AU - Reeve, Shane

AU - Lesk, Arthur

AU - Bottomley, Stephen P.

AU - Rossjohn, Jamie

AU - Whisstock, James C.

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N2 - Serpins fold into a native metastable state and utilize a complex conformational change to inhibit target proteases. An undesirable result of this conformational flexibility is that most inhibitory serpins are heat sensitive, forming inactive polymers at elevated temperatures. However, the prokaryote serpin, thermopin, from Thermobifida fusca is able to function in a heated environment. We have determined the 1.8 Å x-ray crystal structure of thermopin in the native, inhibitory conformation. A structural comparison with the previously determined 1.5 Å structure of cleaved thermopin provides detailed insight into the complex mechanism of conformational change in serpins. Flexibility in the shutter region and electrostatic interactions at the top of the A β-sheet (the breach) involving the C-terminal tail, a unique structural feature of thermopin, are postulated to be important for controlling inhibitory activity and triggering conformational change, respectively, in the native state. Here we have discussed the structural basis of how this serpin reconciles the thermodynamic instability necessary for function with the stability required to withstand elevated temperatures.

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