Recombinant thiopeptides containing noncanonical amino acids

Xiaozhou Luo; Claudio Zambaldo; Tao Liu; Yuhan Zhang; Weimin Xuan; Chen Wang; Sean A. Reed; Peng Yu Yang; Rongsheng E. Wang; Tsotne Javahishvili; Peter G. Schultz; Travis S. Young

doi:10.1073/pnas.1602733113

Recombinant thiopeptides containing noncanonical amino acids

Xiaozhou Luo, Claudio Zambaldo, Tao Liu, Yuhan Zhang, Weimin Xuan, Chen Wang, Sean A. Reed, Peng Yu Yang, Rongsheng E. Wang, Tsotne Javahishvili, Peter G. Schultz, Travis S. Young

Cancer Signaling and Microenvironment (CSM)

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

57 Scopus citations

Abstract

Thiopeptides are a subclass of ribosomally synthesized and posttranslationally modified peptides (RiPPs) with complex molecular architectures and an array of biological activities, including potent antimicrobial activity. Here we report the generation of thiopeptides containing noncanonical amino acids (ncAAs) by introducing orthogonal amber suppressor aminoacyl-tRNA synthetase/tRNA pairs into a thiocillin producer strain of Bacillus cereus. We demonstrate that thiopeptide variants containing ncAAs with bioorthogonal chemical reactivity can be further postbiosynthetically modified with biophysical probes, including fluorophores and photo-cross-linkers. This work allows the site-specific incorporation of ncAAs into thiopeptides to increase their structural diversity and probe their biological activity; similar approaches can likely be applied to other classes of RiPPs.

Original language	English
Pages (from-to)	3615-3620
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	113
Issue number	13
DOIs	https://doi.org/10.1073/pnas.1602733113
State	Published - Mar 29 2016

Keywords

Amino Acid Substitution
Amino Acids/chemistry
Bacillus cereus/genetics
Molecular Structure
Mutagenesis, Site-Directed
Peptides/chemistry
Protein Engineering
Protein Processing, Post-Translational
Recombinant Proteins/chemistry
Tandem Mass Spectrometry

Access to Document

10.1073/pnas.1602733113

Cite this

@article{6217090cf6714fcfa2567f599cfff108,

title = "Recombinant thiopeptides containing noncanonical amino acids",

abstract = "Thiopeptides are a subclass of ribosomally synthesized and posttranslationally modified peptides (RiPPs) with complex molecular architectures and an array of biological activities, including potent antimicrobial activity. Here we report the generation of thiopeptides containing noncanonical amino acids (ncAAs) by introducing orthogonal amber suppressor aminoacyl-tRNA synthetase/tRNA pairs into a thiocillin producer strain of Bacillus cereus. We demonstrate that thiopeptide variants containing ncAAs with bioorthogonal chemical reactivity can be further postbiosynthetically modified with biophysical probes, including fluorophores and photo-cross-linkers. This work allows the site-specific incorporation of ncAAs into thiopeptides to increase their structural diversity and probe their biological activity; similar approaches can likely be applied to other classes of RiPPs.",

keywords = "Amino Acid Substitution, Amino Acids/chemistry, Bacillus cereus/genetics, Molecular Structure, Mutagenesis, Site-Directed, Peptides/chemistry, Protein Engineering, Protein Processing, Post-Translational, Recombinant Proteins/chemistry, Tandem Mass Spectrometry",

author = "Xiaozhou Luo and Claudio Zambaldo and Tao Liu and Yuhan Zhang and Weimin Xuan and Chen Wang and Reed, {Sean A.} and Yang, {Peng Yu} and Wang, {Rongsheng E.} and Tsotne Javahishvili and Schultz, {Peter G.} and Young, {Travis S.}",

year = "2016",

month = mar,

day = "29",

doi = "10.1073/pnas.1602733113",

language = "English",

volume = "113",

pages = "3615--3620",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "13",

}

TY - JOUR

T1 - Recombinant thiopeptides containing noncanonical amino acids

AU - Luo, Xiaozhou

AU - Zambaldo, Claudio

AU - Liu, Tao

AU - Zhang, Yuhan

AU - Xuan, Weimin

AU - Wang, Chen

AU - Reed, Sean A.

AU - Yang, Peng Yu

AU - Wang, Rongsheng E.

AU - Javahishvili, Tsotne

AU - Schultz, Peter G.

AU - Young, Travis S.

PY - 2016/3/29

Y1 - 2016/3/29

N2 - Thiopeptides are a subclass of ribosomally synthesized and posttranslationally modified peptides (RiPPs) with complex molecular architectures and an array of biological activities, including potent antimicrobial activity. Here we report the generation of thiopeptides containing noncanonical amino acids (ncAAs) by introducing orthogonal amber suppressor aminoacyl-tRNA synthetase/tRNA pairs into a thiocillin producer strain of Bacillus cereus. We demonstrate that thiopeptide variants containing ncAAs with bioorthogonal chemical reactivity can be further postbiosynthetically modified with biophysical probes, including fluorophores and photo-cross-linkers. This work allows the site-specific incorporation of ncAAs into thiopeptides to increase their structural diversity and probe their biological activity; similar approaches can likely be applied to other classes of RiPPs.

AB - Thiopeptides are a subclass of ribosomally synthesized and posttranslationally modified peptides (RiPPs) with complex molecular architectures and an array of biological activities, including potent antimicrobial activity. Here we report the generation of thiopeptides containing noncanonical amino acids (ncAAs) by introducing orthogonal amber suppressor aminoacyl-tRNA synthetase/tRNA pairs into a thiocillin producer strain of Bacillus cereus. We demonstrate that thiopeptide variants containing ncAAs with bioorthogonal chemical reactivity can be further postbiosynthetically modified with biophysical probes, including fluorophores and photo-cross-linkers. This work allows the site-specific incorporation of ncAAs into thiopeptides to increase their structural diversity and probe their biological activity; similar approaches can likely be applied to other classes of RiPPs.

KW - Amino Acid Substitution

KW - Amino Acids/chemistry

KW - Bacillus cereus/genetics

KW - Molecular Structure

KW - Mutagenesis, Site-Directed

KW - Peptides/chemistry

KW - Protein Engineering

KW - Protein Processing, Post-Translational

KW - Recombinant Proteins/chemistry

KW - Tandem Mass Spectrometry

UR - http://www.scopus.com/inward/record.url?scp=84962124725&partnerID=8YFLogxK

U2 - 10.1073/pnas.1602733113

DO - 10.1073/pnas.1602733113

M3 - Article

C2 - 26976568

SN - 0027-8424

VL - 113

SP - 3615

EP - 3620

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 13

ER -

Recombinant thiopeptides containing noncanonical amino acids

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this