Rapid Chemical Synthesis of Serine Protease Inhibitor Kazal-type 13 (SPINK13) Glycoform by a Combined Method with Glycan Insertion Strategy and Fast-Flow Fmoc SPPS.
SPINK
glycan
glycoprotein
synthesis
thioacid
Journal
Chemistry (Weinheim an der Bergstrasse, Germany)
ISSN: 1521-3765
Titre abrégé: Chemistry
Pays: Germany
ID NLM: 9513783
Informations de publication
Date de publication:
26 Jul 2023
26 Jul 2023
Historique:
received:
28
02
2023
medline:
27
7
2023
pubmed:
9
6
2023
entrez:
9
6
2023
Statut:
ppublish
Résumé
Serine protease inhibitor Kazal type 13 (SPINK13) is a secreted protein that has been recently studied as a therapeutic drug and an interesting biomarker for cancer cells. Although SPINK13 has a consensus sequence (Pro-Asn-Val-Thr) for N-glycosylation, the existence of N-glycosylation and its functions are still unclear. In addition to this, the preparation of glycosylated SPINK 13 has not been examined by both the cell expression method and chemical synthesis. Herein we report the chemical synthesis of the scarce N-glycosylated form of SPINK13 by a rapid synthetic method combined with the chemical glycan insertion strategy and a fast-flow SPPS method. Glycosylated asparagine thioacid was designed to chemoselectively be inserted between two peptide segments where is the sterically bulky Pro-Asn(N-glycan)-Val junction by two coupling reactions which consist of diacyl disulfide coupling (DDC) and thioacid capture ligation (TCL). This insertion strategy successfully afforded the full-length polypeptide of SPINK13 within two steps from glycosylated asparagine thioacid. Because the two peptides used for this synthesis were prepared by a fast-flow SPPS, the total synthetic time of glycoprotein was considerably shortened. This synthetic concept enables us to repetitively synthesize a target glycoprotein easily. Folding experiments afforded well-folded structure confirmed by CD and disulfide bond map. Invasion assays of glycosylated SPINK13 and non-glycosylated SPINK13 with pancreatic cancer cells showed that non-glycosylated SPINK-13 was more potent than that of glycosylated SPINK13.
Identifiants
pubmed: 37294165
doi: 10.1002/chem.202300646
doi:
Substances chimiques
Serine Proteinase Inhibitors
0
Asparagine
7006-34-0
stable plasma protein solution
0
Peptides
0
Glycoproteins
0
Polysaccharides
0
Disulfides
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e202300646Subventions
Organisme : Japan Society for the Promotion of Science
ID : 17H01214, 21H05028, 21H04708
Organisme : Japan Society for the Promotion of Science
ID : 20J20649
Organisme : Mizutani Foundation for Glycoscience
ID : 210074
Informations de copyright
© 2023 Wiley-VCH GmbH.
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