Resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant Antarctic fish parvalbumins
Parvalbumins (PVs) from Antarctic notothenioid fishes display a pattern of thermal adaptation that likely reflects evolutionary changes in protein conformational flexibility. We have used ancestral sequence reconstruction and homology modeling to identify two amino acid changes that could potentiall...
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fthighwire:oai:open-archive.highwire.org:jexbio:215/18/3281 2023-05-15T13:33:24+02:00 Resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant Antarctic fish parvalbumins Whittington, A. Carl Moerland, Timothy S. 2012-09-15 00:00:00.0 text/html http://jeb.biologists.org/cgi/content/short/215/18/3281 https://doi.org/10.1242/jeb.070615 en eng Company of Biologists http://jeb.biologists.org/cgi/content/short/215/18/3281 http://dx.doi.org/10.1242/jeb.070615 Copyright (C) 2012, Company of Biologists Research Articles TEXT 2012 fthighwire https://doi.org/10.1242/jeb.070615 2013-05-26T19:10:53Z Parvalbumins (PVs) from Antarctic notothenioid fishes display a pattern of thermal adaptation that likely reflects evolutionary changes in protein conformational flexibility. We have used ancestral sequence reconstruction and homology modeling to identify two amino acid changes that could potentially account for the present thermal sensitivity pattern of Antarctic fish PVs compared with a PV from a theoretical warm-adapted ancestral fish. To test this hypothesis, ancient PVs were resurrected in the lab using PV from the notothenioid Gobionotothen gibberifrons as a platform for introducing mutations comparable to the reconstructed ancestral PV sequences. The wild-type PV (WT) as well as three mutant expression constructs were engineered: lysine 8 to asparagine (K8N), lysine 26 to asparagine (K26N) and a double mutant (DM). Calcium equilibrium dissociation constants ( K d ) versus temperature curves for all mutants were right-shifted, as predicted, relative to that of WT PV. The K d values for the K8N and K26N single mutants were virtually identical at all temperatures and showed an intermediate level of thermal sensitivity. The DM construct displayed a full conversion of thermal sensitivity pattern to that of a PV from a warm/temperate-adapted fish. Additionally, the K d versus temperature curve for the WT construct revealed greater thermal sensitivity compared with the mutant constructs. Measurements of the rates of Ca2+ dissociation ( k off ) showed that all mutants generally had slower k off values than WT at all temperatures. Calculated rates of Ca2+ binding ( k on ) for the K8N and K26N mutants were similar to values for the WT PV at all temperatures. In contrast, the calculated k on values for the DM PV were faster, providing mechanistic insights into the nature of potentially adaptive changes in Ca2+ binding in this PV. The overall results suggest that the current thermal phenotype of Antarctic PVs can be recapitulated by just two amino acid substitutions. Text Antarc* Antarctic HighWire Press (Stanford University) Antarctic Journal of Experimental Biology |
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Research Articles |
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Research Articles Whittington, A. Carl Moerland, Timothy S. Resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant Antarctic fish parvalbumins |
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Research Articles |
description |
Parvalbumins (PVs) from Antarctic notothenioid fishes display a pattern of thermal adaptation that likely reflects evolutionary changes in protein conformational flexibility. We have used ancestral sequence reconstruction and homology modeling to identify two amino acid changes that could potentially account for the present thermal sensitivity pattern of Antarctic fish PVs compared with a PV from a theoretical warm-adapted ancestral fish. To test this hypothesis, ancient PVs were resurrected in the lab using PV from the notothenioid Gobionotothen gibberifrons as a platform for introducing mutations comparable to the reconstructed ancestral PV sequences. The wild-type PV (WT) as well as three mutant expression constructs were engineered: lysine 8 to asparagine (K8N), lysine 26 to asparagine (K26N) and a double mutant (DM). Calcium equilibrium dissociation constants ( K d ) versus temperature curves for all mutants were right-shifted, as predicted, relative to that of WT PV. The K d values for the K8N and K26N single mutants were virtually identical at all temperatures and showed an intermediate level of thermal sensitivity. The DM construct displayed a full conversion of thermal sensitivity pattern to that of a PV from a warm/temperate-adapted fish. Additionally, the K d versus temperature curve for the WT construct revealed greater thermal sensitivity compared with the mutant constructs. Measurements of the rates of Ca2+ dissociation ( k off ) showed that all mutants generally had slower k off values than WT at all temperatures. Calculated rates of Ca2+ binding ( k on ) for the K8N and K26N mutants were similar to values for the WT PV at all temperatures. In contrast, the calculated k on values for the DM PV were faster, providing mechanistic insights into the nature of potentially adaptive changes in Ca2+ binding in this PV. The overall results suggest that the current thermal phenotype of Antarctic PVs can be recapitulated by just two amino acid substitutions. |
format |
Text |
author |
Whittington, A. Carl Moerland, Timothy S. |
author_facet |
Whittington, A. Carl Moerland, Timothy S. |
author_sort |
Whittington, A. Carl |
title |
Resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant Antarctic fish parvalbumins |
title_short |
Resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant Antarctic fish parvalbumins |
title_full |
Resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant Antarctic fish parvalbumins |
title_fullStr |
Resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant Antarctic fish parvalbumins |
title_full_unstemmed |
Resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant Antarctic fish parvalbumins |
title_sort |
resurrecting prehistoric parvalbumins to explore the evolution of thermal compensation in extant antarctic fish parvalbumins |
publisher |
Company of Biologists |
publishDate |
2012 |
url |
http://jeb.biologists.org/cgi/content/short/215/18/3281 https://doi.org/10.1242/jeb.070615 |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
http://jeb.biologists.org/cgi/content/short/215/18/3281 http://dx.doi.org/10.1242/jeb.070615 |
op_rights |
Copyright (C) 2012, Company of Biologists |
op_doi |
https://doi.org/10.1242/jeb.070615 |
container_title |
Journal of Experimental Biology |
_version_ |
1766042048593395712 |