Combining multi-mutant and modular thermodynamic cycles to measure energetic coupling networks in enzyme catalysis

Charles W. Carter, Srinivas Niranj Chandrasekaran, Violetta Weinreb, Li Li, Tishan Williams

Research output: Contribution to journalArticle

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Abstract

We measured and cross-validated the energetics of networks in Bacillus stearothermophilus Tryptophanyl-tRNA synthetase (TrpRS) using both multi-mutant and modular thermodynamic cycles. Multi-dimensional combinatorial mutagenesis showed that four side chains from this "molecular switch" move coordinately with the active-site Mg2+ ion as the active site preorganizes to stabilize the transition state for amino acid activation. A modular thermodynamic cycle consisting of fulllength TrpRS, its Urzyme, and the Urzyme plus each of the two domains deleted in the Urzyme gives similar energetics. These dynamic linkages, although unlikely to stabilize the transition-state directly, consign the active-site preorganization to domain motion, assuring coupled vectorial behavior.

LanguageEnglish (US)
Article number032101
JournalStructural Dynamics
Volume4
Issue number3
DOIs
StatePublished - May 1 2017

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Tryptophan-tRNA Ligase
thermodynamic cycles
Catalysis
catalysis
enzymes
Enzymes
Thermodynamics
Mutagenesis
Bacilli
stearothermophilus
mutagenesis
Amino acids
Bacillus
Chemical activation
Switches
Ions
Amino Acids
linkages
amino acids
switches

ASJC Scopus subject areas

  • Radiation
  • Instrumentation
  • Condensed Matter Physics
  • Spectroscopy

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Combining multi-mutant and modular thermodynamic cycles to measure energetic coupling networks in enzyme catalysis. / Carter, Charles W.; Chandrasekaran, Srinivas Niranj; Weinreb, Violetta; Li, Li; Williams, Tishan.

In: Structural Dynamics, Vol. 4, No. 3, 032101, 01.05.2017.

Research output: Contribution to journalArticle

Carter, Charles W. ; Chandrasekaran, Srinivas Niranj ; Weinreb, Violetta ; Li, Li ; Williams, Tishan. / Combining multi-mutant and modular thermodynamic cycles to measure energetic coupling networks in enzyme catalysis. In: Structural Dynamics. 2017 ; Vol. 4, No. 3.
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