Structural analysis of leader peptide binding enables leader-free cyanobactin processing

Jesko Koehnke; Greg Mann; Andrew F. Bent; Hannes Ludewig; Sally Shirran; Catherine Botting; Tomas Lebl; Wael E. Houssen; Marcel Jaspars; James H. Naismith

doi:10.1038/nchembio.1841

Structural analysis of leader peptide binding enables leader-free cyanobactin processing

Jesko Koehnke, Greg Mann, Andrew F. Bent, Hannes Ludewig, Sally Shirran, Catherine Botting, Tomas Lebl, Wael E. Houssen, Marcel Jaspars, James H. Naismith^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

132 Citations (Scopus)

Abstract

Regioselective modification of amino acids within the context of a peptide is common to a number of biosynthetic pathways, and many of the resulting products have potential as therapeutics. The ATP-dependent enzyme LynD heterocyclizes multiple cysteine residues to thiazolines within a peptide substrate. The enzyme requires the substrate to have a conserved N-terminal leader for full activity. Catalysis is almost insensitive to immediately flanking residues in the substrate, suggesting that -recognition occurs distant from the active site. Nucleotide and peptide substrate co-complex structures of LynD reveal that the substrate leader peptide binds to and extends the beta-sheet of a conserved domain of LynD, whereas catalysis is accomplished in another conserved domain. The spatial segregation of catalysis from recognition combines seemingly contradictory properties of regioselectivity and promiscuity, and it appears to be a conserved strategy in other peptide-modifying enzymes. A variant of LynD that efficiently processes substrates without a leader peptide has been engineered.

Original language	English
Pages (from-to)	558-563
Number of pages	8
Journal	Nature Chemical Biology
Volume	11
Early online date	22 Jun 2015
DOIs	https://doi.org/10.1038/nchembio.1841
Publication status	Published - Aug 2015

Bibliographical note

This work was supported by grants from the European Research Council (no. 339367, J.H.N. and M.J.), the UK Biotechnology and Biological Sciences Research Council (no. BB/K015508/1, J.H.N. and M.J.) and the Wellcome Trust, which funded the purchase of the TripleTOF 5600 mass spectrometer at the Biomedical Sciences Research Complex Mass Spectrometry and Proteomics Facility, University of St. Andrews (grant no. 094476/Z/10/Z), and the MALDI TOF-TOF Analyser (grant no. WT079272AIA). We acknowledge use of the Diamond (beamlines I02 and I24) and ESRF (beamline ID29) synchrotrons. J.H.N. is a Royal Society Wolfson Merit Award Holder and a 1000 talent scholar of the Chinese Government at Sichuan University.

Keywords

NATURAL-PRODUCTS
PROCHLORON-DIDEMNI
BIOSYNTHESIS
DISCOVERY
HETEROCYCLIZATION
CYCLODEHYDRATIONS
MECHANISM
INSIGHTS
PATHWAY
ENZYMES

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Marcel Jaspars
- Coastal Communities
- Engineering, National Decommissioning Centre
- School of Natural & Computing Sciences, Chemistry - Chair in Chemistry
Person: Academic

Cite this

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title = "Structural analysis of leader peptide binding enables leader-free cyanobactin processing",

abstract = "Regioselective modification of amino acids within the context of a peptide is common to a number of biosynthetic pathways, and many of the resulting products have potential as therapeutics. The ATP-dependent enzyme LynD heterocyclizes multiple cysteine residues to thiazolines within a peptide substrate. The enzyme requires the substrate to have a conserved N-terminal leader for full activity. Catalysis is almost insensitive to immediately flanking residues in the substrate, suggesting that -recognition occurs distant from the active site. Nucleotide and peptide substrate co-complex structures of LynD reveal that the substrate leader peptide binds to and extends the beta-sheet of a conserved domain of LynD, whereas catalysis is accomplished in another conserved domain. The spatial segregation of catalysis from recognition combines seemingly contradictory properties of regioselectivity and promiscuity, and it appears to be a conserved strategy in other peptide-modifying enzymes. A variant of LynD that efficiently processes substrates without a leader peptide has been engineered.",

keywords = "NATURAL-PRODUCTS, PROCHLORON-DIDEMNI, BIOSYNTHESIS, DISCOVERY, HETEROCYCLIZATION, CYCLODEHYDRATIONS, MECHANISM, INSIGHTS, PATHWAY, ENZYMES",

author = "Jesko Koehnke and Greg Mann and Bent, {Andrew F.} and Hannes Ludewig and Sally Shirran and Catherine Botting and Tomas Lebl and Houssen, {Wael E.} and Marcel Jaspars and Naismith, {James H.}",

note = "This work was supported by grants from the European Research Council (no. 339367, J.H.N. and M.J.), the UK Biotechnology and Biological Sciences Research Council (no. BB/K015508/1, J.H.N. and M.J.) and the Wellcome Trust, which funded the purchase of the TripleTOF 5600 mass spectrometer at the Biomedical Sciences Research Complex Mass Spectrometry and Proteomics Facility, University of St. Andrews (grant no. 094476/Z/10/Z), and the MALDI TOF-TOF Analyser (grant no. WT079272AIA). We acknowledge use of the Diamond (beamlines I02 and I24) and ESRF (beamline ID29) synchrotrons. J.H.N. is a Royal Society Wolfson Merit Award Holder and a 1000 talent scholar of the Chinese Government at Sichuan University. ",

year = "2015",

month = aug,

doi = "10.1038/nchembio.1841",

language = "English",

volume = "11",

pages = "558--563",

journal = "Nature Chemical Biology",

issn = "1552-4450",

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AU - Koehnke, Jesko

AU - Mann, Greg

AU - Bent, Andrew F.

AU - Ludewig, Hannes

AU - Shirran, Sally

AU - Botting, Catherine

AU - Lebl, Tomas

AU - Houssen, Wael E.

AU - Jaspars, Marcel

AU - Naismith, James H.

N1 - This work was supported by grants from the European Research Council (no. 339367, J.H.N. and M.J.), the UK Biotechnology and Biological Sciences Research Council (no. BB/K015508/1, J.H.N. and M.J.) and the Wellcome Trust, which funded the purchase of the TripleTOF 5600 mass spectrometer at the Biomedical Sciences Research Complex Mass Spectrometry and Proteomics Facility, University of St. Andrews (grant no. 094476/Z/10/Z), and the MALDI TOF-TOF Analyser (grant no. WT079272AIA). We acknowledge use of the Diamond (beamlines I02 and I24) and ESRF (beamline ID29) synchrotrons. J.H.N. is a Royal Society Wolfson Merit Award Holder and a 1000 talent scholar of the Chinese Government at Sichuan University.

PY - 2015/8

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N2 - Regioselective modification of amino acids within the context of a peptide is common to a number of biosynthetic pathways, and many of the resulting products have potential as therapeutics. The ATP-dependent enzyme LynD heterocyclizes multiple cysteine residues to thiazolines within a peptide substrate. The enzyme requires the substrate to have a conserved N-terminal leader for full activity. Catalysis is almost insensitive to immediately flanking residues in the substrate, suggesting that -recognition occurs distant from the active site. Nucleotide and peptide substrate co-complex structures of LynD reveal that the substrate leader peptide binds to and extends the beta-sheet of a conserved domain of LynD, whereas catalysis is accomplished in another conserved domain. The spatial segregation of catalysis from recognition combines seemingly contradictory properties of regioselectivity and promiscuity, and it appears to be a conserved strategy in other peptide-modifying enzymes. A variant of LynD that efficiently processes substrates without a leader peptide has been engineered.

AB - Regioselective modification of amino acids within the context of a peptide is common to a number of biosynthetic pathways, and many of the resulting products have potential as therapeutics. The ATP-dependent enzyme LynD heterocyclizes multiple cysteine residues to thiazolines within a peptide substrate. The enzyme requires the substrate to have a conserved N-terminal leader for full activity. Catalysis is almost insensitive to immediately flanking residues in the substrate, suggesting that -recognition occurs distant from the active site. Nucleotide and peptide substrate co-complex structures of LynD reveal that the substrate leader peptide binds to and extends the beta-sheet of a conserved domain of LynD, whereas catalysis is accomplished in another conserved domain. The spatial segregation of catalysis from recognition combines seemingly contradictory properties of regioselectivity and promiscuity, and it appears to be a conserved strategy in other peptide-modifying enzymes. A variant of LynD that efficiently processes substrates without a leader peptide has been engineered.

KW - NATURAL-PRODUCTS

KW - PROCHLORON-DIDEMNI

KW - BIOSYNTHESIS

KW - DISCOVERY

KW - HETEROCYCLIZATION

KW - CYCLODEHYDRATIONS

KW - MECHANISM

KW - INSIGHTS

KW - PATHWAY

KW - ENZYMES

U2 - 10.1038/nchembio.1841

DO - 10.1038/nchembio.1841

M3 - Article

SN - 1552-4450

VL - 11

SP - 558

EP - 563

JO - Nature Chemical Biology

JF - Nature Chemical Biology

ER -

Structural analysis of leader peptide binding enables leader-free cyanobactin processing

Abstract

Bibliographical note

Keywords

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Marcel Jaspars

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