2024-03-28T13:16:50Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/551522021-12-28T16:20:09Zcom_10261_22com_10261_1col_10261_275
DIGITAL.CSIC
author
Garrido, Francisco
author
Taylor, John C.
author
Alfonso, Carlos
author
Markham, George D.
author
Pajares, María A.
2012-08-28T15:32:22Z
2012-08-28T15:32:22Z
2012-08-28
Amino Acids 42(1): 361-373 (2012)
0939-4451
http://hdl.handle.net/10261/55152
10.1007/s00726-010-0813-y
1438-2199
21132339
The methionine adenosyltransferase from the thermophile Methanococcus jannaschii is fully and irreversibly unfolded in the presence of guanidinium chloride. Unfolding of this dimeric protein is a three-state process in which a dimeric intermediate could be identified. The less stable secondary structural elements of the protein are the C-terminal ends of β-strands E2 and E6, as deduced from the behavior of tyrosine to tryptophan mutants at residues 72 and 170, which are located in the subunit interface. Unraveling of these elements at the monomer interface may soften intersubunit interactions, leading to the observed 85% activity loss. Accumulation of the intermediate was associated with maintenance of residual activity, an increase in the elution volume of the protein upon gel filtration and a decrease in the sedimentation coefficient. Elimination of the remaining enzymatic activity occurred in conjunction with a 50% reduction in helicity and fluorescence alterations illustrating a transient burial of tryptophans at β-strands E2, E3 and E9. The available 3D-model predicted that these β-strands are involved in the central and N-terminal domains of the monomer structure. Severe perturbation of this area of the monomer-monomer interface may destroy the remaining intermolecular interactions, thus leading to dissociation and aggregation. Finally, transition to the denatured state includes completion of the changes detected in the microenvironments around tryptophans included at α-helixes H5 and H6, the loops connecting H5 to E8 and E9, β-strands E3 and E12.
eng
openAccess
Denaturation
Stability
Methionine adenosyltransferase
Structural basis for the stability of a thermophilic methionine adenosyltransferase against guanidinium chloride
artículo
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URL
https://digital.csic.es/bitstream/10261/55152/1/Structural%20basis.pdf
File
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Structural basis.pdf
URL
https://digital.csic.es/bitstream/10261/55152/4/Structural%20basis.pdf.txt
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text/plain
Structural basis.pdf.txt