Semi-microbiological synthesis of an active lysinoalanine-bridged analog of glucagon-like-peptide-1

Autor: Rick Rink, Louwe de Vries, Gert N. Moll, Marcel P. de Vries, Anneke Kuipers, Tjibbe Bosma
Přispěvatelé: Molecular Genetics
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Threonine
Physiology
Lysine
Lysinoalanine
POSTTRANSLATIONAL MODIFICATION
Biochemistry
Dehydroalanine
Substrate Specificity
Lanthipeptide
chemistry.chemical_compound
Endocrinology
AMIDE
Glucagon-Like Peptide 1
Serine
Nisin
Alanine
GLUCAGON-LIKE PEPTIDE-1
biology
digestive
oral
and skin physiology
SUBSTRATE-SPECIFICITY
Lactococcus lactis
Glucagon like peptide-1
Signal peptide
endocrine system
Stereochemistry
030106 microbiology
LANTIBIOTIC NISIN
03 medical and health sciences
Cellular and Molecular Neuroscience
BETA-CELLS
Bacterial Proteins
Amino Acid Sequence
LACTOCOCCUS-LACTIS
Hydro-Lyases
RECEPTOR
Membrane Proteins
Membrane Transport Proteins
IN-VITRO
biology.organism_classification
030104 developmental biology
chemistry
Dehydratase
GLP-1
Protein Processing
Post-Translational
Alpha helix
SEC Translocation Channels
Zdroj: Peptides, 91, 33-39. ELSEVIER SCIENCE INC
ISSN: 0196-9781
Popis: Some modified glucagon-like-peptide-1 (GLP-1) analogs are highly important for treating type 2 diabetes. Here we investigated whether GLP-1 analogs expressed in Lactococcus lactis could be substrates for modification and export by the nisin dehydratase and transporter enzyme. Subsequently we introduced a lysinoalanine by coupling a formed dehydroalanine with a lysine and investigated the structure and activity of the formed lysinoalanine-bridged GLP-1 analog. Our data show: (i) GLP-1 fused to the nisin leader peptide is very well exported via the nisin transporter NisT, (ii) production of leader-GLP-1 via NisT is higher than via the SECsystem, (iii) leader-GLP-1 exported via NisT was more efficiently dehydrated by the nisin dehydratase NisB than when exported via the SEC system, (iv) individual serines and threonines in GLP-1 are dehydrated by NisB to a significantly different extent, (v) an introduced Ser30 is well dehydrated and can be coupled to Lys34 to form a lysinoalanine-bridged GLP-1 analog, (vi) a lysinoalanine(30-34) variant's conformation shifts in the presence of 25% trifluoroethanol towards a higher alpha helix content than observed for wild type GLP-1 under identical condition, (vii) a lysinoalanine(30-34) GLP-1 variant has retained significant activity. Taken together the data extend knowledge on the substrate specificities of NisT and NisB and their combined activity relative to export via the Sec system, and demonstrate that introducing a lysinoalanine bridge is an option for modifying therapeutic peptides. (C) 2017 Elsevier Inc. All rights reserved.
Databáze: OpenAIRE
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