This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Tsujimura, M. Articles by Maeda, Y. Search for Related Content PubMed PubMed Citation Articles by Tsujimura, M. Articles by Maeda, Y.

 Previous Article  |  Next Article 

Clinical and Diagnostic Laboratory Immunology, March 2001, p. 454-459, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.454-459.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Detection of Serum Thermolabile -2 Macroglycoprotein (Hakata Antigen) by Enzyme-Linked Immunosorbent Assay Using Polysaccharide Produced by Aerococcus viridans

Mitsushi Tsujimura,¹ Chuzo Ishida,¹ Yasuko Sagara,¹ Takashi Miyazaki,¹ Koichi Murakami,² Hiroshi Shiraki,^1,* Kazuo Okochi,¹ and Yoshiaki Maeda¹

Fukuoka Red Cross Blood Center, Chikushino, Fukuoka 818-8588,¹ and Pathology and Bacteriology Division, Fukuoka Institute of Health and Environmental Sciences, Dazaifu, Fukuoka 818-0135,² Japan

Received 29 June 2000/Returned for modification 19 October 2000/Accepted 11 December 2000

Although a serum thermolabile -2 macroglycoprotein (TMG) may play a role in host defense as a lectin, little is known of its related physiological functions, mainly due to a lack of appropriate methods for tracing the functions of TMG. We identified a polysaccharide from Aerococcus viridans, PSA, which reacts with TMG, and based on this finding, we developed an enzyme-linked immunosorbent assay to trace the functions of TMG. Using ethanol precipitation and DEAE-Sepharose and Sephacryl S-400 column chromatographies, we isolated PSA from cultured medium of A. viridans, and it exhibited specific binding against TMG in blood samples. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the isolated PSA showed ladder bands that implied the existence of repeating units composed of D-glucose, N-acetyl-D-glucosamine, D-mannose, and D-xylose, as confirmed by gas chromatography-mass spectrometry. SDS-PAGE and immunochemical analysis, using rabbit anti-TMG antibody, showed that PSA specifically binds solely to intact serum TMG but not to TMG heated at 56°C for 30 min, a condition under which antigenicity is lost. TMG in serum samples bound to PSA in a dose-dependent manner, and this binding was clearly suppressed by addition of PSA. These observations indicate that PSA is a useful adsorbent to TMG and can be used to develop appropriate methods for tracing the functions of TMG.

---

^* Corresponding author. Mailing address: Fukuoka Red Cross Blood Center, 1-2-1 Kamikoga, Chikushino, Fukuoka 818-8588, Japan. Phone: 81-92-921-1400. Fax: 81-92-921-0799. E-mail: fbc.shiraki{at}fukuoka.emailne.jp.

---

Clinical and Diagnostic Laboratory Immunology, March 2001, p. 454-459, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.454-459.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

• Krarup, A., Thiel, S., Hansen, A., Fujita, T., Jensenius, J. C. (2004). L-ficolin Is a Pattern Recognition Molecule Specific for Acetyl Groups. J. Biol. Chem. 279: 47513-47519 [Abstract] [Full Text]  
• Lynch, N. J., Roscher, S., Hartung, T., Morath, S., Matsushita, M., Maennel, D. N., Kuraya, M., Fujita, T., Schwaeble, W. J. (2004). L-Ficolin Specifically Binds to Lipoteichoic Acid, a Cell Wall Constituent of Gram-Positive Bacteria, and Activates the Lectin Pathway of Complement. J. Immunol. 172: 1198-1202 [Abstract] [Full Text]  
• Matsushita, M., Kuraya, M., Hamasaki, N., Tsujimura, M., Shiraki, H., Fujita, T. (2002). Activation of the Lectin Complement Pathway by H-Ficolin (Hakata Antigen). J. Immunol. 168: 3502-3506 [Abstract] [Full Text]