Identification of mesothelioma-specific sialylated epitope recognized with monoclonal antibody SKM9-2 in a mucin-like membrane protein HEG1

Rieko Matsuura, Hiroyuki Kaji, Azusa Tomioka, Takashi Sato, Hisashi Narimatsu, Yasuhiro Moriwaki, Hidemi Misawa, Kohzoh Imai, Shoutaro Tsuji

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Abstract

The anti-mesothelioma mAb SKM9-2 recognizes the sialylated protein HEG homolog 1 (HEG1). HEG1 is a 400 kDa mucin-like membrane protein found on mesothelioma. SKM9-2 can detect mesothelioma more specifically and sensitively than other antibodies against current mesothelioma markers; therefore, SKM9-2 would be likely useful for the precise detection and diagnosis of malignant mesothelioma. In the present study, we investigated the epitope of SKM9-2. We analyzed the binding of SKM9-2 to truncated HEG1 and candidate epitope-fused glycosylphosphatidylinositol-anchor proteins. The epitope of SKM9-2 was identified as an O-glycosylated region, 893-SKSPSLVSLPT-903, in HEG1. An alanine scanning assay of the epitope showed that SKM9-2 bound to a simple epitope in HEG1, and the SKxPSxVS sequence within the epitope was essential for SKM9-2 recognition. Mass spectrometry analysis and lectin binding analysis of soluble epitope peptides indicated that the SKM9-2 epitope, in which Ser897 was not glycosylated, contained two disialylated core 1 O-linked glycan-modified serine residues, Ser893 and Ser900. Neuraminidase treatment analysis also confirmed that the epitope in mesothelioma cells contained a similar glycan modification. The specific detection of mesothelioma with SKM9-2 can thus be performed by the recognition of sialylated glycan modification in the specific region of HEG1.

Original languageEnglish
Article number14251
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 2018 Dec 1

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Mesothelioma
Mucins
Epitopes
Membrane Proteins
Monoclonal Antibodies
Polysaccharides
Glycosylphosphatidylinositols
Neuraminidase
Lectins
Alanine
Serine
Mass Spectrometry
Proteins
Peptides
Antibodies

ASJC Scopus subject areas

  • General

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Identification of mesothelioma-specific sialylated epitope recognized with monoclonal antibody SKM9-2 in a mucin-like membrane protein HEG1. / Matsuura, Rieko; Kaji, Hiroyuki; Tomioka, Azusa; Sato, Takashi; Narimatsu, Hisashi; Moriwaki, Yasuhiro; Misawa, Hidemi; Imai, Kohzoh; Tsuji, Shoutaro.

In: Scientific Reports, Vol. 8, No. 1, 14251, 01.12.2018.

Research output: Contribution to journalArticle

Matsuura, Rieko ; Kaji, Hiroyuki ; Tomioka, Azusa ; Sato, Takashi ; Narimatsu, Hisashi ; Moriwaki, Yasuhiro ; Misawa, Hidemi ; Imai, Kohzoh ; Tsuji, Shoutaro. / Identification of mesothelioma-specific sialylated epitope recognized with monoclonal antibody SKM9-2 in a mucin-like membrane protein HEG1. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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