Positive charges promote the recognition of proteins by the chaperone SlyD from Escherichia coli

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Autoren

  • Daniel Lindemeier
  • Wenke Graubner
  • Denise Mehner-Breitfeld
  • Miroslav Malesevic
  • Thomas Brüser

Organisationseinheiten

Externe Organisationen

  • Max-Planck-Forschungsstelle für Enzymologie der Proteinfaltung
Forschungs-netzwerk anzeigen

Details

OriginalspracheEnglisch
Aufsatznummere0305823
FachzeitschriftPLOS ONE
Jahrgang19
Ausgabenummer6
PublikationsstatusVeröffentlicht - 25 Juni 2024

Abstract

SlyD is a widely-occurring prokaryotic FKBP-family prolyl isomerase with an additional chaperone domain. Often, such as in Escherichia coli, a third domain is found at its C-terminus that binds nickel and provides it for nickel-enzyme biogenesis. SlyD has been found to bind signal peptides of proteins that are translocated by the Tat pathway, a system for the transport of folded proteins across membranes. Using peptide arrays to analyze these signal peptide interactions, we found that SlyD interacted only with positively charged peptides, with a preference for arginines over lysines, and large hydrophobic residues enhanced binding. Especially a twin-arginine motif was recognized, a pair of highly conserved arginines adjacent to a stretch of hydrophobic residues. Using isothermal titration calorimetry (ITC) with purified SlyD and a signal peptide-containing model Tat substrate, we could show that the wild type twin-arginine signal peptide was bound with higher affinity than an RR>KK mutated variant, confirming that positive charges are recognized by SlyD, with a preference of arginines over lysines. The specific role of negative charges of the chaperone domain surface and of hydrophobic residues in the chaperone active site was further analyzed by ITC of mutated SlyD variants. Our data show that the supposed key hydrophobic residues of the active site are indeed crucial for binding, and that binding is influenced by negative charges on the chaperone domain. Recognition of positive charges is likely achieved by a large negatively charged surface region of the chaperone domain, which is highly conserved although individual positions are variable.

Schlagwörter

    Tat system, Chaperone, Proteininteraktionen, Tat Transport, Escherichia coli

ASJC Scopus Sachgebiete

Zitieren

Positive charges promote the recognition of proteins by the chaperone SlyD from Escherichia coli. / Lindemeier, Daniel; Graubner, Wenke; Mehner-Breitfeld, Denise et al.
in: PLOS ONE, Jahrgang 19, Nr. 6, e0305823, 25.06.2024.

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Lindemeier D, Graubner W, Mehner-Breitfeld D, Malesevic M, Brüser T. Positive charges promote the recognition of proteins by the chaperone SlyD from Escherichia coli. PLOS ONE. 2024 Jun 25;19(6):e0305823. doi: 10.1371/journal.pone.0305823
Lindemeier, Daniel ; Graubner, Wenke ; Mehner-Breitfeld, Denise et al. / Positive charges promote the recognition of proteins by the chaperone SlyD from Escherichia coli. in: PLOS ONE. 2024 ; Jahrgang 19, Nr. 6.
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abstract = "SlyD is a widely-occurring prokaryotic FKBP-family prolyl isomerase with an additional chaperone domain. Often, such as in Escherichia coli, a third domain is found at its C-terminus that binds nickel and provides it for nickel-enzyme biogenesis. SlyD has been found to bind signal peptides of proteins that are translocated by the Tat pathway, a system for the transport of folded proteins across membranes. Using peptide arrays to analyze these signal peptide interactions, we found that SlyD interacted only with positively charged peptides, with a preference for arginines over lysines, and large hydrophobic residues enhanced binding. Especially a twin-arginine motif was recognized, a pair of highly conserved arginines adjacent to a stretch of hydrophobic residues. Using isothermal titration calorimetry (ITC) with purified SlyD and a signal peptide-containing model Tat substrate, we could show that the wild type twin-arginine signal peptide was bound with higher affinity than an RR>KK mutated variant, confirming that positive charges are recognized by SlyD, with a preference of arginines over lysines. The specific role of negative charges of the chaperone domain surface and of hydrophobic residues in the chaperone active site was further analyzed by ITC of mutated SlyD variants. Our data show that the supposed key hydrophobic residues of the active site are indeed crucial for binding, and that binding is influenced by negative charges on the chaperone domain. Recognition of positive charges is likely achieved by a large negatively charged surface region of the chaperone domain, which is highly conserved although individual positions are variable.",
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T1 - Positive charges promote the recognition of proteins by the chaperone SlyD from Escherichia coli

AU - Lindemeier, Daniel

AU - Graubner, Wenke

AU - Mehner-Breitfeld, Denise

AU - Malesevic, Miroslav

AU - Brüser, Thomas

N1 - Publisher Copyright: Copyright: © 2024 Lindemeier et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PY - 2024/6/25

Y1 - 2024/6/25

N2 - SlyD is a widely-occurring prokaryotic FKBP-family prolyl isomerase with an additional chaperone domain. Often, such as in Escherichia coli, a third domain is found at its C-terminus that binds nickel and provides it for nickel-enzyme biogenesis. SlyD has been found to bind signal peptides of proteins that are translocated by the Tat pathway, a system for the transport of folded proteins across membranes. Using peptide arrays to analyze these signal peptide interactions, we found that SlyD interacted only with positively charged peptides, with a preference for arginines over lysines, and large hydrophobic residues enhanced binding. Especially a twin-arginine motif was recognized, a pair of highly conserved arginines adjacent to a stretch of hydrophobic residues. Using isothermal titration calorimetry (ITC) with purified SlyD and a signal peptide-containing model Tat substrate, we could show that the wild type twin-arginine signal peptide was bound with higher affinity than an RR>KK mutated variant, confirming that positive charges are recognized by SlyD, with a preference of arginines over lysines. The specific role of negative charges of the chaperone domain surface and of hydrophobic residues in the chaperone active site was further analyzed by ITC of mutated SlyD variants. Our data show that the supposed key hydrophobic residues of the active site are indeed crucial for binding, and that binding is influenced by negative charges on the chaperone domain. Recognition of positive charges is likely achieved by a large negatively charged surface region of the chaperone domain, which is highly conserved although individual positions are variable.

AB - SlyD is a widely-occurring prokaryotic FKBP-family prolyl isomerase with an additional chaperone domain. Often, such as in Escherichia coli, a third domain is found at its C-terminus that binds nickel and provides it for nickel-enzyme biogenesis. SlyD has been found to bind signal peptides of proteins that are translocated by the Tat pathway, a system for the transport of folded proteins across membranes. Using peptide arrays to analyze these signal peptide interactions, we found that SlyD interacted only with positively charged peptides, with a preference for arginines over lysines, and large hydrophobic residues enhanced binding. Especially a twin-arginine motif was recognized, a pair of highly conserved arginines adjacent to a stretch of hydrophobic residues. Using isothermal titration calorimetry (ITC) with purified SlyD and a signal peptide-containing model Tat substrate, we could show that the wild type twin-arginine signal peptide was bound with higher affinity than an RR>KK mutated variant, confirming that positive charges are recognized by SlyD, with a preference of arginines over lysines. The specific role of negative charges of the chaperone domain surface and of hydrophobic residues in the chaperone active site was further analyzed by ITC of mutated SlyD variants. Our data show that the supposed key hydrophobic residues of the active site are indeed crucial for binding, and that binding is influenced by negative charges on the chaperone domain. Recognition of positive charges is likely achieved by a large negatively charged surface region of the chaperone domain, which is highly conserved although individual positions are variable.

KW - Tat system

KW - Chaperone

KW - Proteininteraktionen

KW - Tat Transport

KW - Escherichia coli

KW - Tat system

KW - Chaperones

KW - Protein interactions

KW - Twin arginine translocation

KW - Escherichia coli

UR - http://www.scopus.com/inward/record.url?scp=85196938119&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0305823

DO - 10.1371/journal.pone.0305823

M3 - Article

VL - 19

JO - PLOS ONE

JF - PLOS ONE

SN - 1932-6203

IS - 6

M1 - e0305823

ER -

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