Electrophoretic characterization of heat-stable squamous cell carcinoma antigen

✍ Scribed by Shugo Nawata; Yoshinori Suminami; Hiroshi Hirakawa; Akihiro Murakami; Kenji Umayahara; Hidenobu Ogata; Fumitaka Numa; Kazuyuki Nakamura; Hiroshi Kato

• Publisher: John Wiley and Sons
• Year: 2001
• Tongue: English
• Weight: 73 KB
• Volume: 22
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/1522-2683(200109)22:16<3522::aid-elps3522>3.0.co;2-#

✦ Synopsis

Electrophoretic characterization of heat-stable squamous cell carcinoma antigen

The aim of this study was to investigate the heat stability of squamous cell carcinoma (SCC) antigen, a tumor-associated serine proteinase inhibitor (serpin), in tumor tissue extract by electrophoretic methods. After heat treatment at 707C for 2 h, the tumor tissue extract showed a single main protein band of 45 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) which reacted with a monoclonal antibody specific for SCC antigen. The heat-stable SCC antigen was separated by two-dimensional electrophoresis (2-DE) into four spots with pI 6.4-5.9 and M r 44 500-45 000 of SCC antigen-1. Furthermore, the SCC antigen-1 still showed its inhibitory activity against a cysteine proteinase, papain, by gelatin zymography. These results suggest that heat treatment of protein sample at 707C for 2 h may be a useful method for a partial purification of SCC antigen-1 which can inhibit lysosomal cysteine proteinases such as cathepsin L, S, and K.