High yield expression and purification of full-length Neurotensin with pyroglutamate modification.

Animals Neurotensin / genetics Pyrrolidonecarboxylic Acid Peptides / chemistry Glutamine Mammals
Isotope labeling Neurotensin Pyroglutamate SUMO

Journal

Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496

Informations de publication

Date de publication:
Apr 2023
Historique:
received: 30 11 2022
revised: 17 12 2022
accepted: 21 12 2022
pubmed: 28 12 2022
medline: 25 1 2023
entrez: 27 12 2022
Statut: ppublish

Résumé

Neurotensin (NT) is a 13-residue endogenous peptide found in mammals, with neurotransmission and hormonal roles in the central nervous system and gastrointestinal tract, respectively. The first residue of NT is a pyroglutamate (pGlu) that makes the expression and purification of large amounts of NT with native modification challenging. Here, we describe a simple and efficient procedure for expression and purification of large amounts of NT based on using the small ubiquitin-like modifier (SUMO) as a fusion partner and subsequent enzymatic conversion of the N-terminal glutamine to pGlu. Yields of 13 mg/L and 8 mg/L of pure peptide were obtained from expression in rich and minimal media, respectively. The method is adaptable to expression and purification of proteins and peptides with pGlu modification in a wide range of eukaryotic and prokaryotic expression hosts.

Identifiants

DOI: 10.1016/j.pep.2022.106227 PMID: 36574939
pubmed: 36574939
pii: S1046-5928(22)00184-X
doi: 10.1016/j.pep.2022.106227
pii:
doi:

Substances chimiques

Neurotensin 39379-15-2
Pyrrolidonecarboxylic Acid SZB83O1W42
Peptides 0
Glutamine 0RH81L854J

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

106227

Informations de copyright

Copyright © 2022 Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of competing interest The authors declare no conflict of interest.

Auteurs

Kazem Asadollahi (K)

Department of Biochemistry and Pharmacology, University of Melbourne, Parkville, VIC, 3052, Australia; Bio21 Molecular Sciences and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3052, Australia; The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, VIC, 3052, Australia.

Katherine Huang (K)

Department of Biochemistry and Pharmacology, University of Melbourne, Parkville, VIC, 3052, Australia; Bio21 Molecular Sciences and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3052, Australia.

Fei Yan (F)

Department of Biochemistry and Pharmacology, University of Melbourne, Parkville, VIC, 3052, Australia; Bio21 Molecular Sciences and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3052, Australia.

Lazarus A de Zhang (LA)

Department of Biochemistry and Pharmacology, University of Melbourne, Parkville, VIC, 3052, Australia; The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, VIC, 3052, Australia; Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC, 3052, Australia.

Daniel J Scott (DJ)

Department of Biochemistry and Pharmacology, University of Melbourne, Parkville, VIC, 3052, Australia; The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, VIC, 3052, Australia.

Paul R Gooley (PR)

Department of Biochemistry and Pharmacology, University of Melbourne, Parkville, VIC, 3052, Australia; Bio21 Molecular Sciences and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3052, Australia. Electronic address: prg@unimelb.edu.au.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › High yield expression and purification of...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines de tissu nerveux › Neuropeptides › Neurotensine › High yield expression and purification of...
questionsmedicales.fr › Acides aminés, peptides et protéines › Acides aminés › Acides aminés cycliques › Imino-acides › Acide pidolique › High yield expression and purification of...
questionsmedicales.fr › Acides aminés, peptides et protéines › Peptides › High yield expression and purification of...
questionsmedicales.fr › Acides aminés, peptides et protéines › Acides aminés › Acides aminés neutres › Glutamine › High yield expression and purification of...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › High yield expression and purification of...