Biomolecular Click Reactions Using a Minimal pH-Activated Catcher/Tag Pair for Producing Native-Sized Spider-Silk Proteins

Ruxia Fan; Johanna Hakanpää; Karoliina Elfving; Helena Taberman; Markus B. Linder; A. Sesilja Aranko

doi:10.1002/anie.202216371

Biomolecular Click Reactions Using a Minimal pH-Activated Catcher/Tag Pair for Producing Native-Sized Spider-Silk Proteins

Ruxia Fan, Johanna Hakanpää, Karoliina Elfving, Helena Taberman, Markus B. Linder^*, A. Sesilja Aranko^*

^*Corresponding author for this work

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

10 Citations (Scopus)

102 Downloads (Pure)

Abstract

A type of protein/peptide pair known as Catcher/Tag pair spontaneously forms an intermolecular isopeptide bond which can be applied for biomolecular click reactions. Covalent protein conjugation using Catcher/Tag pairs has turned out to be a valuable tool in biotechnology and biomedicines, but it is essential to increase the current toolbox of orthogonal Catcher/Tag pairs to expand the range of applications further, for example, for controlled multiple-fragment ligation. We report here the engineering of novel Catcher/Tag pairs for protein ligation, aided by a crystal structure of a minimal CnaB domain from Lactobacillus plantarum. We show that a newly engineered pair, called SilkCatcher/Tag enables efficient pH-inducible protein ligation in addition to being compatible with the widely used SpyCatcher/Tag pair. Finally, we demonstrate the use of the SilkCatcher/Tag pair in the production of native-sized highly repetitive spider-silk-like proteins with >90 % purity, which is not possible by traditional recombinant production methods.

Original language	English
Article number	e202216371
Number of pages	10
Journal	Angewandte Chemie - International Edition
Volume	62
Issue number	11
Early online date	7 Feb 2023
DOIs	https://doi.org/10.1002/anie.202216371
Publication status	Published - 6 Mar 2023
MoE publication type	A1 Journal article-refereed

Keywords

Catcher/Tag
Protein Engineering
Protein Ligation
Protein Modifications
Spider Silk

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10.1002/anie.202216371Licence: CC BY

CHEM_Fan_etal_ Biomolecular Click Reactions Using a Minimal pH‐Activated Catcher Tag Pair for ProducingFinal published version, 4.63 MBLicence: CC BY

Cite this

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title = "Biomolecular Click Reactions Using a Minimal pH-Activated Catcher/Tag Pair for Producing Native-Sized Spider-Silk Proteins",

abstract = "A type of protein/peptide pair known as Catcher/Tag pair spontaneously forms an intermolecular isopeptide bond which can be applied for biomolecular click reactions. Covalent protein conjugation using Catcher/Tag pairs has turned out to be a valuable tool in biotechnology and biomedicines, but it is essential to increase the current toolbox of orthogonal Catcher/Tag pairs to expand the range of applications further, for example, for controlled multiple-fragment ligation. We report here the engineering of novel Catcher/Tag pairs for protein ligation, aided by a crystal structure of a minimal CnaB domain from Lactobacillus plantarum. We show that a newly engineered pair, called SilkCatcher/Tag enables efficient pH-inducible protein ligation in addition to being compatible with the widely used SpyCatcher/Tag pair. Finally, we demonstrate the use of the SilkCatcher/Tag pair in the production of native-sized highly repetitive spider-silk-like proteins with >90 % purity, which is not possible by traditional recombinant production methods.",

keywords = "Catcher/Tag, Protein Engineering, Protein Ligation, Protein Modifications, Spider Silk",

author = "Ruxia Fan and Johanna Hakanp{\"a}{\"a} and Karoliina Elfving and Helena Taberman and Linder, {Markus B.} and Aranko, {A. Sesilja}",

note = "Funding Information: We thank Eva Crosas for her help with the early phases of the structure refinement. Protein crystallization was performed at SPC facility at EMBL Hamburg and the CD spectroscopy and mass spectrometry of the crystallized proteins at the Center for Structural Systems Biology (CSSB, Deutsches Elektronen‐Synchrotron DESY). We acknowledge technical support by the SPC facility at EMBL Hamburg. The synchrotron data was collected at beamline operated by EMBL Hamburg at the PETRA III storage ring (DESY, Hamburg, Germany). This work was supported by the Academy of Finland through its Centres of Excellence Programme Life‐Inspired Hybrid Materials (LIBER, 2022–2029) under project no 346105 and Academy of Finland projects nos. 317395, 308772, and 333238. We are grateful for the support by the FinnCERES Materials Bioeconomy Ecosystem and use of the Bioeconomy Infrastructure at the Aalto University. Publisher Copyright: {\textcopyright} 2023 Wiley-VCH GmbH.",

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doi = "10.1002/anie.202216371",

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AU - Fan, Ruxia

AU - Hakanpää, Johanna

AU - Elfving, Karoliina

AU - Taberman, Helena

AU - Linder, Markus B.

AU - Aranko, A. Sesilja

N1 - Funding Information: We thank Eva Crosas for her help with the early phases of the structure refinement. Protein crystallization was performed at SPC facility at EMBL Hamburg and the CD spectroscopy and mass spectrometry of the crystallized proteins at the Center for Structural Systems Biology (CSSB, Deutsches Elektronen‐Synchrotron DESY). We acknowledge technical support by the SPC facility at EMBL Hamburg. The synchrotron data was collected at beamline operated by EMBL Hamburg at the PETRA III storage ring (DESY, Hamburg, Germany). This work was supported by the Academy of Finland through its Centres of Excellence Programme Life‐Inspired Hybrid Materials (LIBER, 2022–2029) under project no 346105 and Academy of Finland projects nos. 317395, 308772, and 333238. We are grateful for the support by the FinnCERES Materials Bioeconomy Ecosystem and use of the Bioeconomy Infrastructure at the Aalto University. Publisher Copyright: © 2023 Wiley-VCH GmbH.

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N2 - A type of protein/peptide pair known as Catcher/Tag pair spontaneously forms an intermolecular isopeptide bond which can be applied for biomolecular click reactions. Covalent protein conjugation using Catcher/Tag pairs has turned out to be a valuable tool in biotechnology and biomedicines, but it is essential to increase the current toolbox of orthogonal Catcher/Tag pairs to expand the range of applications further, for example, for controlled multiple-fragment ligation. We report here the engineering of novel Catcher/Tag pairs for protein ligation, aided by a crystal structure of a minimal CnaB domain from Lactobacillus plantarum. We show that a newly engineered pair, called SilkCatcher/Tag enables efficient pH-inducible protein ligation in addition to being compatible with the widely used SpyCatcher/Tag pair. Finally, we demonstrate the use of the SilkCatcher/Tag pair in the production of native-sized highly repetitive spider-silk-like proteins with >90 % purity, which is not possible by traditional recombinant production methods.

AB - A type of protein/peptide pair known as Catcher/Tag pair spontaneously forms an intermolecular isopeptide bond which can be applied for biomolecular click reactions. Covalent protein conjugation using Catcher/Tag pairs has turned out to be a valuable tool in biotechnology and biomedicines, but it is essential to increase the current toolbox of orthogonal Catcher/Tag pairs to expand the range of applications further, for example, for controlled multiple-fragment ligation. We report here the engineering of novel Catcher/Tag pairs for protein ligation, aided by a crystal structure of a minimal CnaB domain from Lactobacillus plantarum. We show that a newly engineered pair, called SilkCatcher/Tag enables efficient pH-inducible protein ligation in addition to being compatible with the widely used SpyCatcher/Tag pair. Finally, we demonstrate the use of the SilkCatcher/Tag pair in the production of native-sized highly repetitive spider-silk-like proteins with >90 % purity, which is not possible by traditional recombinant production methods.

KW - Catcher/Tag

KW - Protein Engineering

KW - Protein Ligation

KW - Protein Modifications

KW - Spider Silk

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UR - https://doi.org/10.1101/2022.10.21.513168

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DO - 10.1002/anie.202216371

M3 - Article

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SN - 1433-7851

VL - 62

JO - Angewandte Chemie - International Edition

JF - Angewandte Chemie - International Edition

IS - 11

M1 - e202216371

ER -

Biomolecular Click Reactions Using a Minimal pH-Activated Catcher/Tag Pair for Producing Native-Sized Spider-Silk Proteins

Abstract

Keywords

Access to Document

Other files and links

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FinnCERES: Competence Center for the Materials Bioeconomy: A Flagship for our Sustainable Future

LIBER Linder: Life-like hybrid materials

CrossSilk: Durable and functional biomaterials from crosslinked non-canonical silk and cellulose composite

Bioeconomy Research Infrastructure

Cite this

Biomolecular Click Reactions Using a Minimal pH-Activated Catcher/Tag Pair for Producing Native-Sized Spider-Silk Proteins

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Projects

FinnCERES: Competence Center for the Materials Bioeconomy: A Flagship for our Sustainable Future

LIBER Linder: Life-like hybrid materials

CrossSilk: Durable and functional biomaterials from crosslinked non-canonical silk and cellulose composite

Equipment

Bioeconomy Research Infrastructure

Cite this