NMR solution structure of SlyD from Escherichia coli

Spatial Separation of Prolyl Isomerase and Chaperone Function

verfasst von: Ulrich Weininger, Caroline Haupt, Kristian Schweimer, Wenke Graubner, Michael Kovermann, Thomas Brüser, Christian Scholz, Peter Schaarschmidt, Gabriel Zoldak, Franz X. Schmid, Jochen Balbach
Abstract: SlyD (sensitive to lysis D) is a putative folding helper from the bacterial cytosol and harbors prolyl isomerase and chaperone activities. We determined the solution NMR structure of a truncated version of SlyD (1-165) from Escherichia coli (SlyD^*) that lacks the presumably unstructured C-terminal tail. SlyD^* consists of two well-separated domains: the FKBP domain, which harbors the prolyl isomerase activity, and the insert-in-flap (IF) domain, which harbors the chaperone activity. The IF domain is inserted into a loop of the FKBP domain near the prolyl isomerase active site. The NMR structure of SlyD^* showed no distinct orientation of the two domains relative to each other. In the FKBP domain, Tyr68 points into the active site, which might explain the lowered intrinsic prolyl isomerase activity and the much lower FK506 binding affinity of the protein compared with archetype human FKBP12 (human FK506 binding protein with 12 kDa). The thermodynamics and kinetics of substrate binding by SlyD^* were quantified by fluorescence resonance energy transfer. NMR titration experiments revealed that the IF domain recognizes and binds unfolded or partially folded proteins and peptides. Insulin aggregation is markedly slowed by SlyD^* as evidenced by two-dimensional NMR spectroscopy in real time, probably due to SlyD^* binding to denatured insulin. The capacity of the IF domain to establish an initial encounter-collision complex, together with the flexible orientation of the two interacting domains, makes SlyD^* a very powerful catalyst of protein folding.
Externe Organisation(en): Martin-Luther-Universität Halle-Wittenberg
Universität Bayreuth
Roche Diagnostics GmbH
Typ: Artikel
Journal: Journal of Molecular Biology
Band: 387
Seiten: 295-305
Anzahl der Seiten: 11
ISSN: 0022-2836
Publikationsdatum: 27.03.2009
Publikationsstatus: Veröffentlicht
Peer-reviewed: Ja
ASJC Scopus Sachgebiete: Strukturelle Biologie, Molekularbiologie
Elektronische Version(en): https://doi.org/10.1016/j.jmb.2009.01.034 (Zugang: Unbekannt)

BibTeX

@article{c4e4225ceded4dbc8330d192da759b73,
title = "NMR solution structure of SlyD from Escherichia coli: Spatial Separation of Prolyl Isomerase and Chaperone Function",
abstract = "SlyD (sensitive to lysis D) is a putative folding helper from the bacterial cytosol and harbors prolyl isomerase and chaperone activities. We determined the solution NMR structure of a truncated version of SlyD (1-165) from Escherichia coli (SlyD*) that lacks the presumably unstructured C-terminal tail. SlyD* consists of two well-separated domains: the FKBP domain, which harbors the prolyl isomerase activity, and the insert-in-flap (IF) domain, which harbors the chaperone activity. The IF domain is inserted into a loop of the FKBP domain near the prolyl isomerase active site. The NMR structure of SlyD* showed no distinct orientation of the two domains relative to each other. In the FKBP domain, Tyr68 points into the active site, which might explain the lowered intrinsic prolyl isomerase activity and the much lower FK506 binding affinity of the protein compared with archetype human FKBP12 (human FK506 binding protein with 12 kDa). The thermodynamics and kinetics of substrate binding by SlyD* were quantified by fluorescence resonance energy transfer. NMR titration experiments revealed that the IF domain recognizes and binds unfolded or partially folded proteins and peptides. Insulin aggregation is markedly slowed by SlyD* as evidenced by two-dimensional NMR spectroscopy in real time, probably due to SlyD* binding to denatured insulin. The capacity of the IF domain to establish an initial encounter-collision complex, together with the flexible orientation of the two interacting domains, makes SlyD* a very powerful catalyst of protein folding.",
keywords = "chaperone, NMR, prolyl isomerase, protein folding, SlyD",
author = "Ulrich Weininger and Caroline Haupt and Kristian Schweimer and Wenke Graubner and Michael Kovermann and Thomas Br{\"u}ser and Christian Scholz and Peter Schaarschmidt and Gabriel Zoldak and Schmid, {Franz X.} and Jochen Balbach",
note = "Funding Information: This research was supported by grants from the Deutsche Forschungsgemeinschaft (BA 1821/4-1, SCHM 444/19-1 and GRK 1026 “Conformational Transitions in Macromolecular Interactions”) and the European Regional Development Fund (ERDF). G.Z. was supported by a Deutsche Akademische Austauschdienst postdoctoral fellowship in the program “Modern Applications of Biotechnology.” We thank Paul R{\"o}sch for NMR spectrometer time at 700 and 800 MHz and Christian L{\"o}w for very helpful discussions. We also thank Laurence Thirault, Franz Wagner, Sima Hassanzadeh-Makooi and Nicole Amtmann (Roche, Penzberg) for excellent technical assistance and Dr. Martin Humen{\'i}k for matrix-assisted laser desorption/ionization time-of-flight measurements. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",
year = "2009",
month = mar,
day = "27",
doi = "10.1016/j.jmb.2009.01.034",
language = "English",
volume = "387",
pages = "295--305",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "2",
}