Agonist/Antagonist Activity of Oxytocin Variants Obtained from Free Cyclic Peptide Libraries Generated via Amino Acid Substitution

Kinoshita, Remi; Kozaki, Ikko; Shimizu, Kazunori; Shibata, Takahiro; Ochiai, Akihito; Honda, Hiroyuki

doi:10.1021/acsomega.1c04982

Cited by 6 publications

(2 citation statements)

References 31 publications

(53 reference statements)

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“…However, after replacing the three residues with alanine, EC 50 values of these analogs were not necessarily higher compared with analogs with the replacement of the residues less conserved ( Figure 10 ), which is somewhat unexpected. On the other hand, we found that changes in residues within the disulfide bond ring seem to have a greater impact on receptor activity than those residues outside the ring, similar to results obtained in previous work ( Adachi et al, 2017 ; Kinoshita et al, 2021 ). Similar results using bioassay were also obtained for human urotensin II with a disulfide bond ( Labarrere et al, 2003 ).…”

Section: Discussionsupporting

confidence: 91%

Characterization of an Aplysia vasotocin signaling system and actions of posttranslational modifications and individual residues of the ligand on receptor activity

Ding

Guo

et al. 2023

Front. Pharmacol.

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The vasopressin/oxytocin signaling system is present in both protostomes and deuterostomes and plays various physiological roles. Although there were reports for both vasopressin-like peptides and receptors in mollusc Lymnaea and Octopus, no precursor or receptors have been described in mollusc Aplysia. Here, through bioinformatics, molecular and cellular biology, we identified both the precursor and two receptors for Aplysia vasopressin-like peptide, which we named Aplysia vasotocin (apVT). The precursor provides evidence for the exact sequence of apVT, which is identical to conopressin G from cone snail venom, and contains 9 amino acids, with two cysteines at position 1 and 6, similar to nearly all vasopressin-like peptides. Through inositol monophosphate (IP1) accumulation assay, we demonstrated that two of the three putative receptors we cloned from Aplysia cDNA are true receptors for apVT. We named the two receptors as apVTR1 and apVTR2. We then determined the roles of post-translational modifications (PTMs) of apVT, i.e., the disulfide bond between two cysteines and the C-terminal amidation on receptor activity. Both the disulfide bond and amidation were critical for the activation of the two receptors. Cross-activity with conopressin S, annetocin from an annelid, and vertebrate oxytocin showed that although all three ligands can activate both receptors, the potency of these peptides differed depending on their residue variations from apVT. We, therefore, tested the roles of each residue through alanine substitution and found that each substitution could reduce the potency of the peptide analog, and substitution of the residues within the disulfide bond tended to have a larger impact on receptor activity than the substitution of those outside the bond. Moreover, the two receptors had different sensitivities to the PTMs and single residue substitutions. Thus, we have characterized the Aplysia vasotocin signaling system and showed how the PTMs and individual residues in the ligand contributed to receptor activity.

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Section: Discussionsupporting

confidence: 91%

Characterization of an Aplysia vasotocin signaling system and actions of posttranslational modifications and individual residues of the ligand on receptor activity

Ding

Guo

et al. 2023

Front. Pharmacol.

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“…α-Peptides comprising proteinogenic amino acids usually suffer from poor pharmacological properties, such as low metabolic stability and cell membrane permeability. Specialty peptides, including N-methylated peptides, ,− β-peptides, ,− and cyclic peptides, ,− generally have higher metabolic stability. In particular, it is believed that cyclic N-methylated peptides have better affinity and specificity against their biological targets because of their constrained chemical structure.…”

Section: Microflow Syntheses Of Specialty Peptides Including N-methyl...mentioning

confidence: 99%

Recent Advances in the Solid- and Solution-Phase Synthesis of Peptides and Proteins Using Microflow Technology

Masui

Fuse

2022

Org. Process Res. Dev.

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Peptides have become increasingly important as drugs and drug candidates. In particular, specialty peptides have garnered considerable attention in recent years because of their improved metabolic stability, higher affinity, and biological target selectivity; some of them have the ability to enable cell membrane permeation and oral administration. Despite its long history, peptide synthesis has various limitations, such as high cost, generation of large amounts of waste, and the requirement for hazardous reagents and solvents. Microflow synthesis, wherein the inner diameter of the reaction tube is ≤1 mm, presents several advantages compared with conventional batch synthesis, such as precise control of the reaction time on a short scale, precise reaction temperature control, and facile scale-up with high reproducibility. Microflow technology has been utilized for peptide synthesis since the beginning of the 21st century. The advantages of microflow synthesis enable the use of highly active and unstable chemical species or high-temperature conditions that accelerate peptide synthesis. The combined use of microflow technology, automated synthesis, and online monitoring technologies has emerged in recent years. This approach not only improved the synthetic efficiency but also afforded large amounts of reliable data that can be used to train machine learning models. This review summarizes the solidand solution-phase syntheses of α-peptides and specialty peptides, including N-methylated peptides, β-peptides, and cyclic peptides, reported mainly after 2017. The recently reported mesoflow peptide syntheses (inner diameter of the reaction channels > 1 mm), the microflow automated syntheses, and in-line analysis are covered in this review.

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Discovery of fibroblast growth factor 2‐derived peptides for enhancing mice skeletal muscle satellite cell proliferation

Fujii,

Kinoshita,

Akiyama

et al. 2024

Biotechnology Journal

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Skeletal muscle satellite cells (SCs) are essential for muscle regeneration. Their proliferation and differentiation are influenced by fibroblast growth factor (FGF)‐2. In this study, we screened for FGF‐2‐derived peptides that promote SC proliferation. Utilizing photocleavable peptide array technology, a library of 7‐residue peptides was synthesized, and its effect on SC proliferation was examined using a mixture of five peptides. The results showed that peptides 1–5 (136%), 21–25 (136%), 26–30 (141%), 31–35 (159%), 71–75 (135%), 76–80 (144%), and 126–130 (137%) significantly increased SC proliferation. Further experiments revealed that peptide 33, CKNGGFF, enhanced SC proliferation. Furthermore, its extended form, peptide 33‐13, CKNGGFFLRIHPD, promoted SC proliferation and increased the percentage of Pax7‐positive cells, indicating that SCs were maintained in an undifferentiated state. The addition of FGF‐2 and peptide 33‐13 further induced cell proliferation but did not increase the percentage of Pax7‐positive cells. A proliferation assay using an FGF receptor (FGFR) inhibitor suggested that peptide 33‐13 acts through the FGFR‐mediated and other pathways. Although further research is necessary to explore the mechanisms of action of these peptides and their potential for in vivo and in vitro use, the high sequence conservation of peptides 33 and 33‐13 in FGF‐2 across multiple species suggests their broad application prospects in biomedical engineering and biotechnology.

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Agonist/Antagonist Activity of Oxytocin Variants Obtained from Free Cyclic Peptide Libraries Generated via Amino Acid Substitution

Cited by 6 publications

References 31 publications

Characterization of an Aplysia vasotocin signaling system and actions of posttranslational modifications and individual residues of the ligand on receptor activity

Characterization of an Aplysia vasotocin signaling system and actions of posttranslational modifications and individual residues of the ligand on receptor activity

Recent Advances in the Solid- and Solution-Phase Synthesis of Peptides and Proteins Using Microflow Technology

Discovery of fibroblast growth factor 2‐derived peptides for enhancing mice skeletal muscle satellite cell proliferation

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