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@ARTICLE{Palm:448261,
author = {Palm, Gottfried Julius and Buchholz, Ina and Werten,
Sebastiaan and Girbardt, Britta and Berndt, Leona and
Delcea, Mihaela and Hinrichs, Winfried},
title = {{T}hermodynamics, cooperativity and stability of the
tetracycline repressor ({T}et{R}) upon tetracycline binding},
journal = {Biochimica et biophysica acta / Proteins and proteomics},
volume = {1868},
number = {6},
issn = {1570-9639},
address = {Amsterdam [u.a.]},
reportid = {PUBDB-2020-03359},
pages = {140404},
year = {2020},
abstract = {Allosteric regulation of the Tet repressor (TetR) homodimer
relies on tetracycline binding that abolishes the affinity
for the DNA operator. Previously, interpretation of circular
dichroism data called for unfolding of the α-helical
DNA-binding domains in absence of binding to DNA or
tetracycline. Our small angle X-ray scattering of TetR(D) in
solution contradicts this unfolding as a physiological
process. Instead, in the core domain crystal structures
analyses show increased immobilisation of helix α9 and two
C-terminal turns of helix α8 upon tetracycline binding.
Tetracycline complexes of TetR(D) and four single-site
alanine variants were characterised by isothermal titration
calorimetry, fluorescence titration, X-ray crystal
structures, and melting curves. Five crystal structures
confirm that Thr103 is a key residue for the allosteric
events of induction, with the T103A variant lacking
induction by any tetracycline. The T103A variant shows
anti-cooperative inducer binding, and a melting curve of the
tetracycline complex different to TetR(D) and other
variants. For the N82A variant inducer binding is clearly
anti-cooperative but triggers the induced conformation.},
cin = {EMBL-User},
ddc = {570},
cid = {I:(DE-H253)EMBL-User-20120814},
pnm = {6G3 - PETRA III (POF3-622)},
pid = {G:(DE-HGF)POF3-6G3},
experiment = {EXP:(DE-H253)P-P12-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:32114262},
UT = {WOS:000527000700004},
doi = {10.1016/j.bbapap.2020.140404},
url = {https://bib-pubdb1.desy.de/record/448261},
}