Peptide macrocyclization by a bifunctional endoprotease

Kalia Bernath-Levin, C. Nelson, A.G. Elliott, A.S. Jayasena, Harvey Millar, D.J. Craik, Josh Mylne

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

© 2015 Elsevier Ltd. All rights reserved. Proteases usually cleave peptides, but under some conditions, they can ligate them. Seeds of the common sunflower contain the 14-residue, backbone-macrocyclic peptide sunflower trypsin inhibitor 1 (SFTI-1) whose maturation from its precursor has a genetic requirement for asparaginyl endopeptidase (AEP). To provide more direct evidence, we developed an in situ assay and used 18O-water to demonstrate that SFTI-1 is excised and simultaneously macrocyclized from its linear precursor. The reaction is inefficient in situ, but a newfound breakdown pathway can mask this inefficiency by reducing the internal disulfide bridge of any acyclic-SFTI to thiols before degrading it. To confirm AEP can directly perform the excision/ligation, we produced several recombinant plant AEPs in E. coli, and one from jack bean could catalyze both a typical cleavage reaction and cleavage-dependent, intramolecular transpeptidation to create SFTI-1. We propose that the evolution of ligating endoproteases enables plants like sunflower and jack bean to stabilize bioactive peptides.
Original languageEnglish
Pages (from-to)571-582
JournalChemistry and Biology
Volume22
Issue number5
DOIs
Publication statusPublished - May 2015

Fingerprint

asparaginylendopeptidase
Jacks
Helianthus
Peptides
Masks
Sulfhydryl Compounds
Disulfides
Escherichia coli
Ligation
Seed
Assays
Seeds
Peptide Hydrolases
Water
sunflower SFTI-1 peptide

Cite this

Bernath-Levin, Kalia ; Nelson, C. ; Elliott, A.G. ; Jayasena, A.S. ; Millar, Harvey ; Craik, D.J. ; Mylne, Josh. / Peptide macrocyclization by a bifunctional endoprotease. In: Chemistry and Biology. 2015 ; Vol. 22, No. 5. pp. 571-582.
@article{60230af2324b43618b74bf72879957ac,
title = "Peptide macrocyclization by a bifunctional endoprotease",
abstract = "{\circledC} 2015 Elsevier Ltd. All rights reserved. Proteases usually cleave peptides, but under some conditions, they can ligate them. Seeds of the common sunflower contain the 14-residue, backbone-macrocyclic peptide sunflower trypsin inhibitor 1 (SFTI-1) whose maturation from its precursor has a genetic requirement for asparaginyl endopeptidase (AEP). To provide more direct evidence, we developed an in situ assay and used 18O-water to demonstrate that SFTI-1 is excised and simultaneously macrocyclized from its linear precursor. The reaction is inefficient in situ, but a newfound breakdown pathway can mask this inefficiency by reducing the internal disulfide bridge of any acyclic-SFTI to thiols before degrading it. To confirm AEP can directly perform the excision/ligation, we produced several recombinant plant AEPs in E. coli, and one from jack bean could catalyze both a typical cleavage reaction and cleavage-dependent, intramolecular transpeptidation to create SFTI-1. We propose that the evolution of ligating endoproteases enables plants like sunflower and jack bean to stabilize bioactive peptides.",
author = "Kalia Bernath-Levin and C. Nelson and A.G. Elliott and A.S. Jayasena and Harvey Millar and D.J. Craik and Josh Mylne",
year = "2015",
month = "5",
doi = "10.1016/j.chembiol.2015.04.010",
language = "English",
volume = "22",
pages = "571--582",
journal = "Chemistry & Biology",
issn = "1074-5521",
publisher = "Cell Press",
number = "5",

}

Peptide macrocyclization by a bifunctional endoprotease. / Bernath-Levin, Kalia; Nelson, C.; Elliott, A.G.; Jayasena, A.S.; Millar, Harvey; Craik, D.J.; Mylne, Josh.

In: Chemistry and Biology, Vol. 22, No. 5, 05.2015, p. 571-582.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Peptide macrocyclization by a bifunctional endoprotease

AU - Bernath-Levin, Kalia

AU - Nelson, C.

AU - Elliott, A.G.

AU - Jayasena, A.S.

AU - Millar, Harvey

AU - Craik, D.J.

AU - Mylne, Josh

PY - 2015/5

Y1 - 2015/5

N2 - © 2015 Elsevier Ltd. All rights reserved. Proteases usually cleave peptides, but under some conditions, they can ligate them. Seeds of the common sunflower contain the 14-residue, backbone-macrocyclic peptide sunflower trypsin inhibitor 1 (SFTI-1) whose maturation from its precursor has a genetic requirement for asparaginyl endopeptidase (AEP). To provide more direct evidence, we developed an in situ assay and used 18O-water to demonstrate that SFTI-1 is excised and simultaneously macrocyclized from its linear precursor. The reaction is inefficient in situ, but a newfound breakdown pathway can mask this inefficiency by reducing the internal disulfide bridge of any acyclic-SFTI to thiols before degrading it. To confirm AEP can directly perform the excision/ligation, we produced several recombinant plant AEPs in E. coli, and one from jack bean could catalyze both a typical cleavage reaction and cleavage-dependent, intramolecular transpeptidation to create SFTI-1. We propose that the evolution of ligating endoproteases enables plants like sunflower and jack bean to stabilize bioactive peptides.

AB - © 2015 Elsevier Ltd. All rights reserved. Proteases usually cleave peptides, but under some conditions, they can ligate them. Seeds of the common sunflower contain the 14-residue, backbone-macrocyclic peptide sunflower trypsin inhibitor 1 (SFTI-1) whose maturation from its precursor has a genetic requirement for asparaginyl endopeptidase (AEP). To provide more direct evidence, we developed an in situ assay and used 18O-water to demonstrate that SFTI-1 is excised and simultaneously macrocyclized from its linear precursor. The reaction is inefficient in situ, but a newfound breakdown pathway can mask this inefficiency by reducing the internal disulfide bridge of any acyclic-SFTI to thiols before degrading it. To confirm AEP can directly perform the excision/ligation, we produced several recombinant plant AEPs in E. coli, and one from jack bean could catalyze both a typical cleavage reaction and cleavage-dependent, intramolecular transpeptidation to create SFTI-1. We propose that the evolution of ligating endoproteases enables plants like sunflower and jack bean to stabilize bioactive peptides.

U2 - 10.1016/j.chembiol.2015.04.010

DO - 10.1016/j.chembiol.2015.04.010

M3 - Article

VL - 22

SP - 571

EP - 582

JO - Chemistry & Biology

JF - Chemistry & Biology

SN - 1074-5521

IS - 5

ER -