Crystal structure of halogenase PltA from the pyoluteorin biosynthetic pathway

Allan H. Pang; Sylvie Garneau-Tsodikova; Oleg V. Tsodikov

doi:10.1016/j.jsb.2015.09.013

Crystal structure of halogenase PltA from the pyoluteorin biosynthetic pathway

Allan H. Pang, Sylvie Garneau-Tsodikova, Oleg V. Tsodikov

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

24 Scopus citations

Abstract

Pyoluteorin is an antifungal agent composed of a 4,5-dichlorinated pyrrole group linked to a resorcinol moiety. The pyoluteorin biosynthetic gene cluster in Pseudomonas fluorescens Pf-5 encodes the halogenase PltA, which has been previously demonstrated to perform both chlorinations in vitro. PltA selectively accepts as a substrate a pyrrole moiety covalently tethered to a nonribosomal peptide thiolation domain PltL (pyrrolyl- S-PltL) for FAD-dependent di-chlorination, yielding 4,5-dichloropyrrolyl- S-PltL. We report a 2.75 Å-resolution crystal structure of PltA in complex with FAD and chloride. PltA is a dimeric enzyme, containing a flavin-binding fold conserved in flavin-dependent halogenases and monooxygenases, and an additional unique helical region at the C-terminus. This C-terminal region blocks a putative substrate-binding cleft, suggesting that a conformational change involving repositioning of this region is necessary to allow binding of the pyrrolyl- S-PltL substrate for its dichlorination by PltA.

Original language	English
Pages (from-to)	349-357
Number of pages	9
Journal	Journal of Structural Biology
Volume	192
Issue number	3
DOIs	https://doi.org/10.1016/j.jsb.2015.09.013
State	Published - Dec 2015

Bibliographical note

Funding Information:
This work was supported by an NSF Career Award MCB-1149427 (to S.G.-T.), a Grant (to O.V.T.) as part of the National Center for Advancing Translational Sciences ( UL1TR000117 ), and startup funds from the College of Pharmacy at the University of Kentucky (to S.G.-T. and O.V.T.). We thank the staff of sector 22 (SER-CAT) of the Advanced Photon Source at the Argonne National Laboratories for their assistance with the remote X-ray diffraction data collection.

Publisher Copyright:
© 2015 Elsevier Inc.

Keywords

Chlorination
FAD
Flavoprotein
Halogenation
Natural product

ASJC Scopus subject areas

Structural Biology

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10.1016/j.jsb.2015.09.013

Cite this

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title = "Crystal structure of halogenase PltA from the pyoluteorin biosynthetic pathway",

abstract = "Pyoluteorin is an antifungal agent composed of a 4,5-dichlorinated pyrrole group linked to a resorcinol moiety. The pyoluteorin biosynthetic gene cluster in Pseudomonas fluorescens Pf-5 encodes the halogenase PltA, which has been previously demonstrated to perform both chlorinations in vitro. PltA selectively accepts as a substrate a pyrrole moiety covalently tethered to a nonribosomal peptide thiolation domain PltL (pyrrolyl- S-PltL) for FAD-dependent di-chlorination, yielding 4,5-dichloropyrrolyl- S-PltL. We report a 2.75 {\AA}-resolution crystal structure of PltA in complex with FAD and chloride. PltA is a dimeric enzyme, containing a flavin-binding fold conserved in flavin-dependent halogenases and monooxygenases, and an additional unique helical region at the C-terminus. This C-terminal region blocks a putative substrate-binding cleft, suggesting that a conformational change involving repositioning of this region is necessary to allow binding of the pyrrolyl- S-PltL substrate for its dichlorination by PltA.",

keywords = "Chlorination, FAD, Flavoprotein, Halogenation, Natural product",

author = "Pang, {Allan H.} and Sylvie Garneau-Tsodikova and Tsodikov, {Oleg V.}",

note = "Funding Information: This work was supported by an NSF Career Award MCB-1149427 (to S.G.-T.), a Grant (to O.V.T.) as part of the National Center for Advancing Translational Sciences ( UL1TR000117 ), and startup funds from the College of Pharmacy at the University of Kentucky (to S.G.-T. and O.V.T.). We thank the staff of sector 22 (SER-CAT) of the Advanced Photon Source at the Argonne National Laboratories for their assistance with the remote X-ray diffraction data collection. Publisher Copyright: {\textcopyright} 2015 Elsevier Inc.",

year = "2015",

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language = "English",

volume = "192",

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AU - Pang, Allan H.

AU - Garneau-Tsodikova, Sylvie

AU - Tsodikov, Oleg V.

N1 - Funding Information: This work was supported by an NSF Career Award MCB-1149427 (to S.G.-T.), a Grant (to O.V.T.) as part of the National Center for Advancing Translational Sciences ( UL1TR000117 ), and startup funds from the College of Pharmacy at the University of Kentucky (to S.G.-T. and O.V.T.). We thank the staff of sector 22 (SER-CAT) of the Advanced Photon Source at the Argonne National Laboratories for their assistance with the remote X-ray diffraction data collection. Publisher Copyright: © 2015 Elsevier Inc.

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Y1 - 2015/12

N2 - Pyoluteorin is an antifungal agent composed of a 4,5-dichlorinated pyrrole group linked to a resorcinol moiety. The pyoluteorin biosynthetic gene cluster in Pseudomonas fluorescens Pf-5 encodes the halogenase PltA, which has been previously demonstrated to perform both chlorinations in vitro. PltA selectively accepts as a substrate a pyrrole moiety covalently tethered to a nonribosomal peptide thiolation domain PltL (pyrrolyl- S-PltL) for FAD-dependent di-chlorination, yielding 4,5-dichloropyrrolyl- S-PltL. We report a 2.75 Å-resolution crystal structure of PltA in complex with FAD and chloride. PltA is a dimeric enzyme, containing a flavin-binding fold conserved in flavin-dependent halogenases and monooxygenases, and an additional unique helical region at the C-terminus. This C-terminal region blocks a putative substrate-binding cleft, suggesting that a conformational change involving repositioning of this region is necessary to allow binding of the pyrrolyl- S-PltL substrate for its dichlorination by PltA.

AB - Pyoluteorin is an antifungal agent composed of a 4,5-dichlorinated pyrrole group linked to a resorcinol moiety. The pyoluteorin biosynthetic gene cluster in Pseudomonas fluorescens Pf-5 encodes the halogenase PltA, which has been previously demonstrated to perform both chlorinations in vitro. PltA selectively accepts as a substrate a pyrrole moiety covalently tethered to a nonribosomal peptide thiolation domain PltL (pyrrolyl- S-PltL) for FAD-dependent di-chlorination, yielding 4,5-dichloropyrrolyl- S-PltL. We report a 2.75 Å-resolution crystal structure of PltA in complex with FAD and chloride. PltA is a dimeric enzyme, containing a flavin-binding fold conserved in flavin-dependent halogenases and monooxygenases, and an additional unique helical region at the C-terminus. This C-terminal region blocks a putative substrate-binding cleft, suggesting that a conformational change involving repositioning of this region is necessary to allow binding of the pyrrolyl- S-PltL substrate for its dichlorination by PltA.

KW - Chlorination

KW - FAD

KW - Flavoprotein

KW - Halogenation

KW - Natural product

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SP - 349

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Crystal structure of halogenase PltA from the pyoluteorin biosynthetic pathway

Abstract

Bibliographical note

Keywords

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