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@ARTICLE{Yu:619633,
author = {Yu, Miao and Gruzinov, Andrey Yu. and Ruan, Hao and
Scheidt, Tom and Chowdhury, Aritra and Giofrè, Sabrina and
Mohammed, Ahmed S. A. and Caria, Joana and Sauter, Paul F.
and Svergun, Dmitri I. and Lemke, Edward A.},
title = {{A} genetically encoded anomalous {SAXS} ruler to probe the
dimensions of intrinsically disordered proteins},
journal = {Proceedings of the National Academy of Sciences of the
United States of America},
volume = {121},
number = {50},
issn = {0027-8424},
address = {Washington, DC},
publisher = {National Acad. of Sciences},
reportid = {PUBDB-2024-07772},
pages = {e2415220121},
year = {2024},
abstract = {Intrinsically disordered proteins (IDPs) adopt ensembles of
rapidly fluctuating heterogeneous conformations, influencing
their binding capabilities and supramolecular transitions.
The primary conformational descriptors for understanding IDP
ensembles—the radius of gyration (RG), measured by
small-angle X-ray scattering (SAXS), and the root mean
square (rms) end-to-end distance (RE), probed by fluorescent
resonance energy transfer (FRET)—are often reported to
produce inconsistent results regarding IDP expansion as a
function of denaturant concentration in the buffer. This
ongoing debate surrounding the FRET-SAXS discrepancy raises
questions about the overall reliability of either method for
quantitatively studying IDP properties. To address this
discrepancy, we introduce a genetically encoded anomalous
SAXS (ASAXS) ruler, enabling simultaneous and direct
measurements of RG and RE without assuming a specific
structural model. This ruler utilizes a genetically encoded
noncanonical amino acid with two bromine atoms, providing an
anomalous X-ray scattering signal for precise distance
measurements. Through this approach, we experimentally
demonstrate that the ratio between RE and RG varies under
different denaturing conditions, highlighting the intrinsic
properties of IDPs as the primary source of the observed
SAXS-FRET discrepancy rather than shortcomings in either of
the two established methods. The developed genetically
encoded ASAXS ruler emerges as a versatile tool for both
IDPs and folded proteins, providing a unified approach for
obtaining complementary and site-specific conformational
information in scattering experiments, thereby contributing
to a deeper understanding of protein functions.},
cin = {EMBL-User / EMBL},
ddc = {500},
cid = {I:(DE-H253)EMBL-User-20120814 / I:(DE-H253)EMBL-20120731},
pnm = {6G3 - PETRA III (DESY) (POF4-6G3) / DFG project
G:(GEPRIS)432343117 - Eine genetisch kodierte ASAXS Methode
zur Messung der Eigenschaften von Intrinsisch ungeordneten
Proteinen (432343117) / DFG project 464588647 - SFB 1551:
Polymerkonzepte zum Verstehen zellulärer Funktionen
(464588647) / iNEXT-Discovery - Infrastructure for
transnational access and discovery in structural biology
(871037)},
pid = {G:(DE-HGF)POF4-6G3 / G:(GEPRIS)432343117 /
G:(GEPRIS)464588647 / G:(EU-Grant)871037},
experiment = {EXP:(DE-H253)P-P12-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:39642200},
UT = {WOS:001380520800016},
doi = {10.1073/pnas.2415220121},
url = {https://bib-pubdb1.desy.de/record/619633},
}