Bioluminescent immunoassay for serine/threonine protein kinase activity using an aequorin-labeled monoclonal antibody and a synthetic peptide as a substrate.


Journal

Biochemical and biophysical research communications
ISSN: 1090-2104
Titre abrégé: Biochem Biophys Res Commun
Pays: United States
ID NLM: 0372516

Informations de publication

Date de publication:
12 11 2023
Historique:
received: 30 08 2023
accepted: 21 09 2023
medline: 23 10 2023
pubmed: 2 10 2023
entrez: 2 10 2023
Statut: ppublish

Résumé

A bioluminescent immunoassay system was developed to determine serine/threonine protein kinase activity using an aequorin-labeled monoclonal antibody and a synthetic peptide as the substrate. A monoclonal antibody against the synthetic phosphorylated serine peptide (K9P peptide) of histone H3 (19 amino acid residues), referred to as the H3S10P antibody, was chemically conjugated to maleimide-activated aequorin to prepare aequorin-labeled H3S10P (AQ-S-H3S10P). For the serine/threonine kinase assay, a non-phosphorylated serine peptide (K9C peptide) coated on a microplate was incubated with serine/threonine protein kinase in the presence of ATP and Mg

Identifiants

pubmed: 37783115
pii: S0006-291X(23)01098-7
doi: 10.1016/j.bbrc.2023.09.062
pii:
doi:

Substances chimiques

Aequorin 50934-79-7
Antibodies, Monoclonal 0
Peptides 0
Protein Serine-Threonine Kinases EC 2.7.11.1
Protein Kinases EC 2.7.-
Threonine 2ZD004190S

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

180-185

Informations de copyright

Copyright © 2023 Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of competing interest J. Sato and S. Inouye are employees in JNC Corporation. H. Onogi, and N. Nomura are employees in KinoPharma, Inc. M. Hagiwara declares that there are no conflicts of interest.

Auteurs

Jun-Ich Sato (JI)

Yokohama Research Center, JNC Co., 5-1 Okawa, Kanazawa-ku, Yokohama, 236-8605, Japan.

Hiroshi Onogi (H)

KinoPharma, Inc. Kyoto-University Katsura Venture Plaza 1-39 Goryoohara, Nishikyo-ku, Kyoto, 615-8245, Japan. Electronic address: honogi@kinopharma.com.

Namiko Nomura (N)

KinoPharma, Inc. Kyoto-University Katsura Venture Plaza 1-39 Goryoohara, Nishikyo-ku, Kyoto, 615-8245, Japan.

Masatoshi Hagiwara (M)

Department of Anatomy and Developmental Biology, Kyoto University Graduate School of Medicine, Sakyo-Ku, Kyoto, 606-8501, Japan.

Satoshi Inouye (S)

Yokohama Research Center, JNC Co., 5-1 Okawa, Kanazawa-ku, Yokohama, 236-8605, Japan. Electronic address: sinouye@dpc.agu.ac.jp.

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Classifications MeSH