Biosensor-Linked Immunosorbent Assay for the Quantification of Methionine Oxidation in Target Proteins

Hae Min Lee, Dong Wook Choi, Seahyun Kim, Aro Lee, Minseo Kim, Yeon Jin Roh, Young Ho Jo, Hwa Yeon Cho, Ho Jae Lee, Seung Rock Lee, Lionel Tarrago, Vadim N. Gladyshev, Ji Hyung Kim, Byung Cheon Lee

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Methionine oxidation is involved in regulating the protein activity and often leads to protein malfunction. However, tools for quantitative analyses of protein-specific methionine oxidation are currently unavailable. In this work, we developed a biological sensor that quantifies oxidized methionine in the form of methionine-R-sulfoxide in target proteins. The biosensor “tpMetROG” consists of methionine sulfoxide reductase B (MsrB), circularly permuted yellow fluorescent protein (cpYFP), thioredoxin, and protein G. Protein G binds to the constant region of antibodies against target proteins, specifically capturing them. Then, MsrB reduces the oxidized methionine in these proteins, leading to cpYFP fluorescence changes. We assessed this biosensor for quantitative analysis of methionine-R-sulfoxide in various proteins, such as calmodulin, IDLO, LegP, Sacde, and actin. We further developed an immunosorbent assay using the biosensor to quantify methionine oxidation in specific proteins such as calmodulin in animal tissues. The biosensor-linked immunosorbent assay proves to be an indispensable tool for detecting methionine oxidation in a protein-specific manner. This is a versatile tool for studying the redox biology of methionine oxidation in proteins.

Original languageEnglish
Pages (from-to)131-141
Number of pages11
JournalACS Sensors
Volume7
Issue number1
DOIs
Publication statusPublished - 2022 Jan 28

Keywords

  • biosensor
  • immunosorbent assay
  • methionine oxidation
  • methionine sulfoxide reductase B
  • redox

ASJC Scopus subject areas

  • Bioengineering
  • Instrumentation
  • Process Chemistry and Technology
  • Fluid Flow and Transfer Processes

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