An adaptive mutation in adenylate kinase that increases organismal fitness is linked to stability-activity trade-offs

Counago, Rafael, Wilson, Corey J., Pena, Matthew I., Wittung-Stafshede, Pernilla and Shamoo, Yousif (2008) An adaptive mutation in adenylate kinase that increases organismal fitness is linked to stability-activity trade-offs. Protein Engineering Design and Selection, 21 1: 19-27.


Author Counago, Rafael
Wilson, Corey J.
Pena, Matthew I.
Wittung-Stafshede, Pernilla
Shamoo, Yousif
Title An adaptive mutation in adenylate kinase that increases organismal fitness is linked to stability-activity trade-offs
Journal name Protein Engineering Design and Selection  (ERA 2012 Listed)    (ERA 2010 Rank B)   Check publisher's open access policy
Publication date 2008-01
Sub-type Article
DOI 10.1093/protein/gzm072
Volume number 21
Issue number 1
ISSN 1741-0126; 1741-0134
Start page 19
End page 27
Total pages 9
Place of publication Oxford, U.K.
Publisher Oxford University Press
Language eng
Formatted abstract Protein function is a balance between activity and stability. However, the relevance of stability–activity trade-offs for protein evolution and their impact on organismal fitness have been difficult to determine. Previously, we have linked organismal survival at increasing temperatures to adaptive changes to a single protein sequence through allelic replacement of an essential gene, adenylate kinase (adk), in a thermophile. In vivo continuous evolution of the temperature-sensitive thermophile has shown that the first step toward increased organismal fitness is mutation of glutamine-199 to arginine in the mesophilic enzyme (AKsub Q199R). Here, we show that although substitution of Arg-199 did confer a modest increase in stability (0.6 kcal mol−1at 20°C; ΔTm = 3.0°C), it is a large change in the activity profile of the enzyme that is responsible for its exceptional robustness during the earlier experimental evolution study. Kinetic studies of AKsub Q199R show that it has a strong loss of enzymatic activity (>50%) at lower temperatures (20–45°C) and a subsequent increase at elevated temperatures. The stability–activity trade-off observed for AKsub Q199R was linked to the rigidification of the overall structure through stabilization of a polypeptide loop containing Arg-199 that is part of the ATP-binding site of the enzyme. Structural analysis revealed the formation of new ionic interactions facilitated by Arg-199. Our results suggest that stability–activity trade-offs are employed readily as an evolutionary strategy during natural selection to increase organismal fitness.
Keyword Adaptive evolution
Adenylate kinase
Molecular evolution
Thermostability
Trade-offs
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article
Collections: ERA 2012 Admin Only
School of Chemistry and Molecular Biosciences
 
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