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^*

^*Tämän työn vastaava kirjoittaja

Tutkimustuotos: Lehtiartikkeli › Article › Scientific › vertaisarvioitu

10 Sitaatiot (Scopus)

108 Lataukset (Pure)

Abstrakti

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.

Alkuperäiskieli	Englanti
Artikkeli	e202216371
Sivumäärä	10
Julkaisu	Angewandte Chemie - International Edition
Vuosikerta	62
Numero	11
Varhainen verkossa julkaisun päivämäärä	7 helmik. 2023
DOI - pysyväislinkit	https://doi.org/10.1002/anie.202216371
Tila	Julkaistu - 6 maalisk. 2023
OKM-julkaisutyyppi	A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

Pääsy asiakirjaan

10.1002/anie.202216371Lisenssi: CC BY

CHEM_Fan_etal_ Biomolecular Click Reactions Using a Minimal pH‐Activated Catcher Tag Pair for ProducingFinal published version, 4,63 MBLisenssi: CC BY

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2 Aktiivinen
3 Päättynyt

FinnCERES: Competence Center for the Materials Bioeconomy: A Flagship for our Sustainable Future
Naukkarinen, O. (Vastuullinen tutkija)
01/05/2022 → 30/06/2026
Projekti: Academy of Finland: Other research funding
LIBER Linder: Life-like hybrid materials
Linder, M. (Vastuullinen tutkija), Elfving, K. (Projektin jäsen), Lemetti, L. (Projektin jäsen), Tunn, I. (Projektin jäsen), Aspelin, H. (Projektin jäsen), Roas Escalona, N. (Projektin jäsen), Aranko, S. (Projektin jäsen), Malkamäki, M. (Projektin jäsen), Osmekhina, E. (Projektin jäsen) & Fedorov, D. (Projektin jäsen)
01/01/2022 → 31/12/2026
Projekti: Academy of Finland: Other research funding
CrossSilk: Durable and functional biomaterials from crosslinked non-canonical silk and cellulose composite
Aranko, S. (Vastuullinen tutkija), Elfving, K. (Projektin jäsen), Shen, M. (Projektin jäsen), Fan, R. (Projektin jäsen) & Astapov, D. (Projektin jäsen)
01/09/2020 → 31/08/2024
Projekti: Academy of Finland: Other research funding

Biotalousinfrastruktuuri
Seppälä, J. (Manager)
Kemian tekniikan korkeakoulu
Laitteistot/tilat: Facility

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@article{e222e519d5fd4cdab53e2210c02cf2f0,

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.",

year = "2023",

month = mar,

day = "6",

doi = "10.1002/anie.202216371",

language = "English",

volume = "62",

journal = "Angewandte Chemie - International Edition",

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T1 - Biomolecular Click Reactions Using a Minimal pH-Activated Catcher/Tag Pair for Producing Native-Sized Spider-Silk Proteins

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.

PY - 2023/3/6

Y1 - 2023/3/6

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

UR - http://www.scopus.com/inward/record.url?scp=85147357916&partnerID=8YFLogxK

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

U2 - 10.1002/anie.202216371

DO - 10.1002/anie.202216371

M3 - Article

AN - SCOPUS:85147357916

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

Abstrakti

Pääsy asiakirjaan

Muut tiedostot ja linkit

Sormenjälki

Projektit

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

Laitteet

Biotalousinfrastruktuuri

Siteeraa tätä