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...

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Main Authors: Kazumi Fujimoto, Akinori Hayashi, Yoshio Kodera, Tatsuya Saito, Takuya Toki, Akifumi Ogawa, Yuji Kamata, Koji Takano, Hideki Katakami, Masayoshi Shichiri
Format: Article
Language:English
Published: Nature Portfolio 2017-08-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-017-08288-0
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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.
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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
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