Effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens
Despite the presence and abundance of archaea in low-temperature environments, little information is available regarding their physiological and biochemical properties. In order to investigate the adaptation of archaeal proteins to low temperatures, we purified and characterized the elongation facto...
Main Authors: | , |
---|---|
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
2000
|
Subjects: | |
Online Access: | http://hdl.handle.net/1959.4/39580 |
id |
ftunswworks:oai:unsworks.library.unsw.edu.au:1959.4/39580 |
---|---|
record_format |
openpolar |
spelling |
ftunswworks:oai:unsworks.library.unsw.edu.au:1959.4/39580 2023-05-15T13:55:40+02:00 Effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens Thomas, Torsten Cavicchioli, Ricardo 2000 http://hdl.handle.net/1959.4/39580 EN eng http://jb.asm.org/cgi/content/abstract/182/5/1328 http://hdl.handle.net/1959.4/39580 metadata only access http://purl.org/coar/access_right/c_14cb CC BY-NC-ND 3.0 https://creativecommons.org/licenses/by-nc-nd/3.0/au/ CC-BY-NC-ND urn:ISSN:0021-9193 Journal of Bacteriology, 182, 5, 1328-1332 journal article http://purl.org/coar/resource_type/c_6501 2000 ftunswworks 2022-08-09T07:41:50Z Despite the presence and abundance of archaea in low-temperature environments, little information is available regarding their physiological and biochemical properties. In order to investigate the adaptation of archaeal proteins to low temperatures, we purified and characterized the elongation factor 2 (EF-2) protein from the Antarctic methanogen Methanococcoides burtonii, which was expressed in Escherichia coli, and compared it to the recombinant EF-2 protein from a phylogenetically related thermophile, Methanosarcina thermophila. Using differential scanning calorimetry to assess protein stability and enzyme assays for the intrinsic GTPase activity, we identified biochemical and biophysical properties that are characteristic of the cold-adapted protein. This includes a higher activity at low temperatures caused by a decrease of the activation energy necessary for GTP hydrolysis and a decreased activation energy for the irreversible denaturation of the protein, which indicates a less thermostable structure. Comparison of the in vitro properties of the proteins with the temperature-dependent characteristics of growth of the organisms indicates that additional cytoplasmic factors are likely to be important for the complete thermal adaptation of the proteins in vivo. This is the first study to address thermal adaptation of proteins from a free-living, cold-adapted archaeon, and our results indicate that the ability of the Antarctic methanogen to adapt to the cold is likely to involve protein structural changes. Article in Journal/Newspaper Antarc* Antarctic UNSW Sydney (The University of New South Wales): UNSWorks Antarctic The Antarctic |
institution |
Open Polar |
collection |
UNSW Sydney (The University of New South Wales): UNSWorks |
op_collection_id |
ftunswworks |
language |
English |
description |
Despite the presence and abundance of archaea in low-temperature environments, little information is available regarding their physiological and biochemical properties. In order to investigate the adaptation of archaeal proteins to low temperatures, we purified and characterized the elongation factor 2 (EF-2) protein from the Antarctic methanogen Methanococcoides burtonii, which was expressed in Escherichia coli, and compared it to the recombinant EF-2 protein from a phylogenetically related thermophile, Methanosarcina thermophila. Using differential scanning calorimetry to assess protein stability and enzyme assays for the intrinsic GTPase activity, we identified biochemical and biophysical properties that are characteristic of the cold-adapted protein. This includes a higher activity at low temperatures caused by a decrease of the activation energy necessary for GTP hydrolysis and a decreased activation energy for the irreversible denaturation of the protein, which indicates a less thermostable structure. Comparison of the in vitro properties of the proteins with the temperature-dependent characteristics of growth of the organisms indicates that additional cytoplasmic factors are likely to be important for the complete thermal adaptation of the proteins in vivo. This is the first study to address thermal adaptation of proteins from a free-living, cold-adapted archaeon, and our results indicate that the ability of the Antarctic methanogen to adapt to the cold is likely to involve protein structural changes. |
format |
Article in Journal/Newspaper |
author |
Thomas, Torsten Cavicchioli, Ricardo |
spellingShingle |
Thomas, Torsten Cavicchioli, Ricardo Effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens |
author_facet |
Thomas, Torsten Cavicchioli, Ricardo |
author_sort |
Thomas, Torsten |
title |
Effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens |
title_short |
Effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens |
title_full |
Effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens |
title_fullStr |
Effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens |
title_full_unstemmed |
Effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens |
title_sort |
effect of temperature on the stability and activity of the elongation factor 2 proteins from low-temperature adapted and thermophilic methanogens |
publishDate |
2000 |
url |
http://hdl.handle.net/1959.4/39580 |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
urn:ISSN:0021-9193 Journal of Bacteriology, 182, 5, 1328-1332 |
op_relation |
http://jb.asm.org/cgi/content/abstract/182/5/1328 http://hdl.handle.net/1959.4/39580 |
op_rights |
metadata only access http://purl.org/coar/access_right/c_14cb CC BY-NC-ND 3.0 https://creativecommons.org/licenses/by-nc-nd/3.0/au/ |
op_rightsnorm |
CC-BY-NC-ND |
_version_ |
1766262460017278976 |