Work

Clonal Stabilization Reveals a DAO / 3MST ‐Expressing MDCK Subpopulation With Robust d ‐Cysteine‐Mediated H 2 S Production

Year: 2026

Type: article

Abstract: ABSTRACT Hydrogen sulfide (H 2 S) functions as a signaling molecule and cytoprotectant. In many tissues, H 2 S is produced from l ‐cysteine, whereas the kidney also produces H 2 S from d ‐cysteine by the sequential activities of d ‐amino acid oxidase (DAO) and 3‐mercaptopyruvate sulfurtransferase (3MST). The d ‐cysteine pathway produces H 2 S more efficiently than the l ‐cysteine pathway, and d ‐cysteine significantly reduces ischemia–reperfusion injury in the renal cortex compared with l ‐cysteine. However, the specific renal cell types responsible for the d ‐cysteine pathway have not been clearly identified. Here, we show that a subpopulation of MDCK (NBL‐2) cells, a renal epithelial cell line, expresses both DAO and 3MST. MDCK cells initially produced H 2 S from d ‐cysteine, but this activity progressively declined with passage. In contrast, clonal cells expressing DAO and 3MST, established from this subpopulation, maintained stable H 2 S production. The clonal cells exhibited two distinct patterns of DAO expression: one showed increased DAO levels after confluence, whereas the other displayed high DAO expression even before reaching confluence. These findings suggest that the d ‐cysteine pathway operates in epithelial cells and plays distinct roles depending on cellular state and dynamics. Moreover, the established clonal cells may provide insight into the role of d ‐cysteine‐mediated H 2 S production in the kidney.

Source: The FASEB Journal

DOI: 10.1096/fj.202503702rr

Authors: Akari Miyamoto, Haruna Ueno, Hideo Kimura, Tadashi Nakagawa, Norihiro Shibuya

Institution: Sanyo-Onoda City University

Language: English

Topic: Sulfur Compounds in Biology

Subfield: Biochemistry

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $3,740