Identification and quantification of plasma free salusin-β, an endogenous parasympathomimetic peptide
Abstract Salusin-β is an endogenous parasympathomimetic proatherosclerotic peptide. Salusin-β was initially predicted from bioinformatic analyses and later immunologically detected in human biofluids. However, elucidation of salusin-β bioactivity has faced enormous challenges because of its unique p...
Main Authors: | , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Nature Portfolio
2017-08-01
|
Series: | Scientific Reports |
Online Access: | https://doi.org/10.1038/s41598-017-08288-0 |
_version_ | 1818752534045523968 |
---|---|
author | Kazumi Fujimoto Akinori Hayashi Yoshio Kodera Tatsuya Saito Takuya Toki Akifumi Ogawa Yuji Kamata Koji Takano Hideki Katakami Masayoshi Shichiri |
author_facet | Kazumi Fujimoto Akinori Hayashi Yoshio Kodera Tatsuya Saito Takuya Toki Akifumi Ogawa Yuji Kamata Koji Takano Hideki Katakami Masayoshi Shichiri |
author_sort | Kazumi Fujimoto |
collection | DOAJ |
description | Abstract Salusin-β is an endogenous parasympathomimetic proatherosclerotic peptide. Salusin-β was initially predicted from bioinformatic analyses and later immunologically detected in human biofluids. However, elucidation of salusin-β bioactivity has faced enormous challenges because of its unique physicochemical characteristics that cause it to strongly adhere to laboratory apparatus materials. In the strictest sense, the discovery of bioactive peptides is not complete until their exact native sequences have been confirmed in the peripheral circulation. In this study, we determined the plasma molecular form and levels of free salusin-β to determine its pathophysiological significance. Ultra-high-yield enrichment and preseparation of non-tryptic human plasma was followed by LC-MS/MS, and full-length salusin-β and seven different endogenous fragment sequences were identified. We established a new ELISA that specifically detects plasma free salusin-β without cross-reacting with any of its identified endogenous fragments. Free salusin-β levels exhibited a profound early morning nadir and rapidly decreased in response to parasympathetic nervous augmentation. Our technical advance in plasma native peptide analysis successfully identified a hard-to-detect bioactive peptide, salusin-β, together with its formerly unrecognized fragments, and further suggests that conventional immunological measurements of target peptides may not be fully representative. |
first_indexed | 2024-12-18T04:52:59Z |
format | Article |
id | doaj.art-12e669158622491cb39e2ea5cf573602 |
institution | Directory Open Access Journal |
issn | 2045-2322 |
language | English |
last_indexed | 2024-12-18T04:52:59Z |
publishDate | 2017-08-01 |
publisher | Nature Portfolio |
record_format | Article |
series | Scientific Reports |
spelling | doaj.art-12e669158622491cb39e2ea5cf5736022022-12-21T21:20:22ZengNature PortfolioScientific Reports2045-23222017-08-01711910.1038/s41598-017-08288-0Identification and quantification of plasma free salusin-β, an endogenous parasympathomimetic peptideKazumi Fujimoto0Akinori Hayashi1Yoshio Kodera2Tatsuya Saito3Takuya Toki4Akifumi Ogawa5Yuji Kamata6Koji Takano7Hideki Katakami8Masayoshi Shichiri9Department of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-kuDepartment of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-kuDepartment of Physics, Laboratory of Biophysics, Kitasato University School of Science, 1-15-1 Kitasato, Minami-kuDepartment of Physics, Laboratory of Biophysics, Kitasato University School of Science, 1-15-1 Kitasato, Minami-kuDepartment of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-kuDepartment of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-kuDepartment of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-kuDepartment of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-kuTeikyo University Chiba Medical CenterDepartment of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-kuAbstract Salusin-β is an endogenous parasympathomimetic proatherosclerotic peptide. Salusin-β was initially predicted from bioinformatic analyses and later immunologically detected in human biofluids. However, elucidation of salusin-β bioactivity has faced enormous challenges because of its unique physicochemical characteristics that cause it to strongly adhere to laboratory apparatus materials. In the strictest sense, the discovery of bioactive peptides is not complete until their exact native sequences have been confirmed in the peripheral circulation. In this study, we determined the plasma molecular form and levels of free salusin-β to determine its pathophysiological significance. Ultra-high-yield enrichment and preseparation of non-tryptic human plasma was followed by LC-MS/MS, and full-length salusin-β and seven different endogenous fragment sequences were identified. We established a new ELISA that specifically detects plasma free salusin-β without cross-reacting with any of its identified endogenous fragments. Free salusin-β levels exhibited a profound early morning nadir and rapidly decreased in response to parasympathetic nervous augmentation. Our technical advance in plasma native peptide analysis successfully identified a hard-to-detect bioactive peptide, salusin-β, together with its formerly unrecognized fragments, and further suggests that conventional immunological measurements of target peptides may not be fully representative.https://doi.org/10.1038/s41598-017-08288-0 |
spellingShingle | Kazumi Fujimoto Akinori Hayashi Yoshio Kodera Tatsuya Saito Takuya Toki Akifumi Ogawa Yuji Kamata Koji Takano Hideki Katakami Masayoshi Shichiri Identification and quantification of plasma free salusin-β, an endogenous parasympathomimetic peptide Scientific Reports |
title | Identification and quantification of plasma free salusin-β, an endogenous parasympathomimetic peptide |
title_full | Identification and quantification of plasma free salusin-β, an endogenous parasympathomimetic peptide |
title_fullStr | Identification and quantification of plasma free salusin-β, an endogenous parasympathomimetic peptide |
title_full_unstemmed | Identification and quantification of plasma free salusin-β, an endogenous parasympathomimetic peptide |
title_short | Identification and quantification of plasma free salusin-β, an endogenous parasympathomimetic peptide |
title_sort | identification and quantification of plasma free salusin β an endogenous parasympathomimetic peptide |
url | https://doi.org/10.1038/s41598-017-08288-0 |
work_keys_str_mv | AT kazumifujimoto identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT akinorihayashi identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT yoshiokodera identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT tatsuyasaito identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT takuyatoki identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT akifumiogawa identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT yujikamata identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT kojitakano identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT hidekikatakami identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide AT masayoshishichiri identificationandquantificationofplasmafreesalusinbanendogenousparasympathomimeticpeptide |