1. Academic Validation
  2. Effects of salazosulfapyridine on the profile of cell surface proteins, revealed by biotinylation of cell surface proteins and 2-dimentional electrophoresis

Effects of salazosulfapyridine on the profile of cell surface proteins, revealed by biotinylation of cell surface proteins and 2-dimentional electrophoresis

  • Biochim Biophys Acta Proteins Proteom. 2019 Jan;1867(1):47-56. doi: 10.1016/j.bbapap.2018.05.007.
Kazuki Omoteyama 1 Toshiyuki Sato 2 Mitsumi Arito 2 Masaaki Sato 2 Naoya Suematsu 2 Manae S Kurokawa 3 Tomohiro Kato 2
Affiliations

Affiliations

  • 1 Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine. Electronic address: omoteyama@marianna-u.ac.jp.
  • 2 Clinical Proteomics and Molecular Medicine, St. Marianna University Graduate School of Medicine.
  • 3 Disease Biomarker Analysis and Molecular Regulation, St. Marianna University Graduate School of Medicine.
Abstract

Objective: We investigated effects of salazosulfapyridine (SASP) on the protein profile of cell surface (CS)-proteins of SW982, a human synovial sarcoma cell line, using biotinylation of CS-proteins and 2-dimensional fluorescence difference gel electrophoresis (2D-DIGE).

Methods: SW982 cells were treated with SASP and its metabolites, sulfapyridine (SP) and 5-aminosalicylic acid (5ASA). Then the cells were treated with a membrane-impermeable biotinylating reagent. Biotinylated CS-proteins were isolated using NeutrAvidin-bound beads. CS-proteins affected by the drugs were detected by 2D-DIGE and subjected to mass spectrometry.

Results: By the 2D-DIGE analysis, in total 576 spots were detected, 29 out of which showed more than ±1.5-fold different intensity in the SASP-, SP-, and 5ASA-treated cells, compared to non-treated cells (p < 0.05). Interestingly, 7 out of the 29 spots changed their intensity only by SASP and 17 spots changed their intensity only by SP. We identified 9 protein from 15 out of the 29 spots, most of which were evidenced to exist on the cell surface by flow cytometry.

Conclusion: We found novel effects of SASP and its metabolites on SW982 cells by the combination of biotinylation of cell surface proteins and 2D-DIGE analysis. These data would help understanding of anti-rheumatic actions of SASP. Furthermore, the combination would be a useful method for the analysis of CS-proteins in various conditions.

Keywords

Biotinylation; Cell surface protein; Rheumatoid arthritis; Salazosulfapyridine; Two-dimensional electrophoresis.

Figures
Products