Details
| Original language | English |
|---|---|
| Pages (from-to) | 295-305 |
| Number of pages | 11 |
| Journal | Journal of Molecular Biology |
| Volume | 387 |
| Issue number | 2 |
| Publication status | Published - 27 Mar 2009 |
| Externally published | Yes |
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
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Structural Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
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In: Journal of Molecular Biology, Vol. 387, No. 2, 27.03.2009, p. 295-305.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - NMR solution structure of SlyD from Escherichia coli
T2 - Spatial Separation of Prolyl Isomerase and Chaperone Function
AU - Weininger, Ulrich
AU - Haupt, Caroline
AU - Schweimer, Kristian
AU - Graubner, Wenke
AU - Kovermann, Michael
AU - Brüser, Thomas
AU - Scholz, Christian
AU - Schaarschmidt, Peter
AU - Zoldak, Gabriel
AU - Schmid, Franz X.
AU - Balbach, Jochen
N1 - 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 Rösch for NMR spectrometer time at 700 and 800 MHz and Christian Löw 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 Humeník for matrix-assisted laser desorption/ionization time-of-flight measurements. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2009/3/27
Y1 - 2009/3/27
N2 - 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.
AB - 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.
KW - chaperone
KW - NMR
KW - prolyl isomerase
KW - protein folding
KW - SlyD
UR - http://www.scopus.com/inward/record.url?scp=61649089934&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2009.01.034
DO - 10.1016/j.jmb.2009.01.034
M3 - Article
C2 - 19356587
AN - SCOPUS:61649089934
VL - 387
SP - 295
EP - 305
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 2
ER -