title: Identification of mucins by using a method involving a combination of on-membrane chemical deglycosylation and immunostaining.
abstract: The characterization of mucins is critically important for gaining insights into the molecular pathology of diseases, including cancers, as well as for the discovery of biomarkers for disease diagnosis. However, no practical method has yet been reported for identifying mucin proteins. Here, we report a technique for immunological identification of mucins separated by supported molecular matrix electrophoresis (SMME), a recently developed membrane electrophoresis method. The technique involves on-membrane deglycosylation of mucins by using mild periodate oxidation/base-catalyzed elimination, followed by immunostaining with an antibody that specifically recognizes the mucin tandem repeat (TR) peptide. We demonstrated the method's feasibility by using MUC1 derived from 2 cancer cell lines, T47D and HPAF-II. The present method shows potential as an alternative approach for the identification of mucins separated by SMME or blotted from conventional gel electrophoresis, on a PVDF membrane.
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Open Space Laboratory C-2, 1-1-1 Umezono, Tsukuba, Ibaraki 305-8568, Japan.
authors.forename: Yu-ki
authors.initials: YK
authors.lastname: Matsuno
authors.forename: Weijie
authors.initials: W
authors.lastname: Dong
authors.forename: Seiya
authors.initials: S
authors.lastname: Yokoyama
authors.forename: Suguru
authors.initials: S
authors.lastname: Yonezawa
authors.forename: Hisashi
authors.initials: H
authors.lastname: Narimatsu
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kameyama
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D004586:Electrophoresis; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D013194:Staining and Labeling
chemical_list: D018396:Mucin-1
keywords: Antibody; Chemical deglycosylation; Identification; Mucins; On-membrane; SMME; Supported molecular matrix electrophoresis; TFMSA; TR; supported molecular matrix electrophoresis; tandem repeat; trifluoromethanesulfonic acid