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@ARTICLE{Fitter:42202,
author = {Fitter, J. and Haber-Pohlmeier, S.},
title = {{S}tructural {S}tability and {U}nfolding {P}roperties of
{T}hermostable bacterial alpha-amylases: {A} {C}omparative
{S}tudy on {H}omologous {E}nzymes},
journal = {Biochemistry},
volume = {43},
issn = {0006-2960},
address = {Columbus, Ohio},
publisher = {American Chemical Society},
reportid = {PreJuSER-42202},
pages = {9589 - 9599},
year = {2004},
note = {Record converted from VDB: 12.11.2012},
abstract = {In a comparative investigation on two thermostable
alpha-amylases [Bacillus amyloliquefaciens (BAA), T(m) = 86
degrees C and Bacillus licheniformis (BLA), T(m) = 101
degrees C], we studied thermal and guanidine hydrochloride
(GndHCl)-induced unfolding using fluorescence and CD
spectroscopy, as well as dynamic light scattering. Depletion
of calcium from specific ion-binding sites in the protein
structures reduces the melting temperature tremendously for
both alpha-amylases. The reduction is nearly the same for
both enzymes, namely, in the order of 50 degrees C. Thus,
the difference in thermostability between BLA and BAA
(DeltaT(m) approximately 15 degrees C) is related to
intrinsic properties of the respective protein structures
themselves and is not related to the strength of ion
binding. The thermal unfolding of both proteins is
characterized by a full disappearance of secondary structure
elements and by a concurrent expansion of the 3D structure.
GndHCl-induced unfolding also yields a fully vanishing
secondary structure but with more expanded 3D structures.
Both alpha-amylases remain much more compact upon thermal
unfolding as compared to the fully unfolded state induced by
chemical denaturants. Such rather compact thermal unfolded
structures lower the conformational entropy change during
the unfolding transition, which principally can contribute
to an increased thermal stability. Structural flexibilities
of both enzymes, as measured with tryptophan fluorescence
quenching, are almost identical for both enzymes in the
native states, as well as in the unfolded states.
Furthermore, we do not observe any difference in the
temperature dependence of the structural flexibilities
between BLA and BAA. These results indicate that
conformational dynamics on the time scale of our studies
seem not to be related to thermal stability or to thermal
adaptation.},
keywords = {Bacillus: enzymology / Bacterial Proteins: chemistry /
Enzyme Stability / Guanidine: chemistry / Kinetics / Light /
Protein Denaturation / Protein Folding / Scattering,
Radiation / Sequence Homology, Amino Acid / Spectrometry,
Fluorescence / Temperature / Thermodynamics /
alpha-Amylases: chemistry / Bacterial Proteins (NLM
Chemicals) / Guanidine (NLM Chemicals) / alpha-Amylases (NLM
Chemicals) / J (WoSType)},
cin = {IBI-2},
ddc = {570},
cid = {I:(DE-Juel1)VDB58},
pnm = {Neurowissenschaften},
pid = {G:(DE-Juel1)FUEK255},
shelfmark = {Biochemistry $\&$ Molecular Biology},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:15274613},
UT = {WOS:000222965100002},
doi = {10.1021/bi0493362},
url = {https://juser.fz-juelich.de/record/42202},
}