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Article

Sphingomyelin Deacylase, the Enzyme Involved in the Pathogenesis of Atopic Dermatitis, Is Identical to the β-Subunit of Acid Ceramidase

1
Drug Research Division, Dainippon Sumitomo Pharma Co., Ltd., Osaka City, Osaka 554-0022, Japan
2
Department of Dermatology, Tokyo Women’s Medical University, Tokyo 162-8666, Japan
3
Center for Bioscience Research & Education, Utsunomiya University, 350 Mine Utsunomiya, Tochigi 321-8505, Japan
4
National Institutes of Biomedical Innovation, Health and Nutrition, AI Center for Health and Biomedical Research 7-6-8 Asagi Saito Ibaraki-city, Osaka 567-0085, Japan
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Current address: DSP Business Partners Co., Ltd. R&D support Business Unit, R&D Technical Support Service Division, 1-98, Kasugade-naka 3-chome, Konohana-ku, Osaka 554-0022, Japan.
§
Current address: Bioscience Research Laboratory, Sumitomo Chemical Co., Ltd. 1-98, Kasugade-naka 3-chome, Konohana-ku, Osaka 554-0022, Japan.
Int. J. Mol. Sci. 2020, 21(22), 8789; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228789
Received: 26 October 2020 / Revised: 13 November 2020 / Accepted: 16 November 2020 / Published: 20 November 2020
(This article belongs to the Special Issue Molecular Mechanisms of Skin Aging and Atopic Dermatitis)
A ceramide deficiency in the stratum corneum (SC) is an essential etiologic factor for the dry and barrier-disrupted skin of patients with atopic dermatitis (AD). Previously, we reported that sphingomyelin (SM) deacylase, which hydrolyzes SM and glucosylceramide at the acyl site to yield their lysoforms sphingosylphosphorylcholine (SPC) and glucosylsphingosine, respectively, instead of ceramide and/or acylceramide, is over-expressed in AD skin and results in a ceramide deficiency. Although the enzymatic properties of SM deacylase have been clarified, the enzyme itself remains unidentified. In this study, we purified and characterized SM deacylase from rat skin. The activities of SM deacylase and acid ceramidase (aCDase) were measured using SM and ceramide as substrates by tandem mass spectrometry by monitoring the production of SPC and sphingosine, respectively. Levels of SM deacylase activity from various rat organs were higher in the order of skin > lung > heart. By successive chromatography using Phenyl-5PW, Rotofor, SP-Sepharose, Superdex 200 and Shodex RP18-415, SM deacylase was purified to homogeneity with a single band of an apparent molecular mass of 43 kDa with an enrichment of > 14,000-fold. Analysis by MALDI-TOF MS/MS using a protein spot with SM deacylase activity separated by 2D-SDS-PAGE allowed its amino acid sequence to be determined and identified as the β-subunit of aCDase, which consists of α- and β-subunits linked by amino bonds and a single S-S bond. Western blotting of samples treated with 2-mercaptoethanol revealed that, whereas recombinant human aCDase was recognized by antibodies to the α-subunit at ~56 kDa and ~13 kDa and the β-subunit at ~43 kDa, the purified SM deacylase was detectable only by the antibody to the β-subunit at ~43 kDa. Breaking the S-S bond of recombinant human aCDase with dithiothreitol elicited the activity of SM deacylase with ~40 kDa upon gel chromatography. These results provide new insights into the essential role of SM deacylase expressed as an aCDase-degrading β-subunit that evokes the ceramide deficiency in AD skin. View Full-Text
Keywords: atopic dermatitis; ceramide; ceramide deficiency; barrier function; water reservoir faction; stratum corneum; sphingomyelin deacylase; sphingosylphosphorylcholine; acid ceramidase atopic dermatitis; ceramide; ceramide deficiency; barrier function; water reservoir faction; stratum corneum; sphingomyelin deacylase; sphingosylphosphorylcholine; acid ceramidase
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MDPI and ACS Style

Teranishi, Y.; Kuwahara, H.; Ueda, M.; Takemura, T.; Kusumoto, M.; Nakamura, K.; Sakai, J.; Kimura, T.; Furutani, Y.; Kawashima, M.; Imokawa, G.; Nogami-Itoh, M. Sphingomyelin Deacylase, the Enzyme Involved in the Pathogenesis of Atopic Dermatitis, Is Identical to the β-Subunit of Acid Ceramidase. Int. J. Mol. Sci. 2020, 21, 8789. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228789

AMA Style

Teranishi Y, Kuwahara H, Ueda M, Takemura T, Kusumoto M, Nakamura K, Sakai J, Kimura T, Furutani Y, Kawashima M, Imokawa G, Nogami-Itoh M. Sphingomyelin Deacylase, the Enzyme Involved in the Pathogenesis of Atopic Dermatitis, Is Identical to the β-Subunit of Acid Ceramidase. International Journal of Molecular Sciences. 2020; 21(22):8789. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228789

Chicago/Turabian Style

Teranishi, Yasuhiro; Kuwahara, Hiroshi; Ueda, Masaru; Takemura, Tadashi; Kusumoto, Masanori; Nakamura, Keiji; Sakai, Jun; Kimura, Toru; Furutani, Yasuji; Kawashima, Makoto; Imokawa, Genji; Nogami-Itoh, Mari. 2020. "Sphingomyelin Deacylase, the Enzyme Involved in the Pathogenesis of Atopic Dermatitis, Is Identical to the β-Subunit of Acid Ceramidase" Int. J. Mol. Sci. 21, no. 22: 8789. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21228789

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