Volume 3 Supplement 2

21st European Workshop for Rheumatology Research

Open Access

Structure and function of an autoantigen, alpha-enloase

  • S Moscato1,
  • F Bongiorni1,
  • F Pratesi1,
  • M Scavuzzo1,
  • S Bombardieri1 and
  • P Migliorini1
Arthritis Research & Therapy20013(Suppl 2):P073

DOI: 10.1186/ar242

Received: 15 January 2001

Published: 26 January 2001

The glycolytic enzyme 2-phosphoglyceratelyase (alpha-enolase) is an autoantigen in connective tissue disorders, and more frequently in patients with active renal disease. The enzyme has pleiotropic functions: it is also a structural protein, a stress protein induced by hypoxia and it acts as transcription factor in the nucleus. Alpha enolase is encoded by a single copy gene and only one mRNA species is detected. In order to define a structural basis for these different functions, we analyzed the isoelectric point of the enzyme. On a kidney extract fractionated by 2D electrophoresis, a mouse anti-enolase antiserum detects 5 spots of identical molecular weight but differing in pI. Some autoimmune sera react with all the spots, while other recognize only the acidic forms of alpha-enolase. We then analyzed the properties of the membrane form of enolase. Enolase is not a membrane structural protein, but it is strongly associated with the membrane, where it acts as plasminogen receptor. Anti-enolase antibodies purified from autoimmune sera react also with the membrane form of alpha-enolase: by flow cytometry, 7/9 antibody preparations bind in fact U937 cells, a human lymphomonocytoid cell line that expresses high density of plasminogen receptors. To investigate the possible functional role of membrane enolase, we evaluated the ability of monoclonal anti-enolase antibodies to induce cell damage or apoptosis. No monoclonal had a cytotoxic effect on U937 cells or was able to induce apoptosis in the same cell line. We then tested the ability of monoclonal anti-enolase antibodies to induce Ca2+ influx in U937 cells. One out of 4 monoclonal antibodies induced release of Ca2+ from intracellular stores.

In conclusion, alpha-enolase exists as multiple isoforms, probably due to postranslational modifications, which seem to affect recognition by autoantibodies. It is presently unknown whether these modifications are tissue-specific and/or affect membrane expression of the enzyme. A possible link between Ca2+ influx and receptor functions of enolase is currently under investigation.

Authors’ Affiliations

(1)
Clinical Immunology Unit, Department of Internal Medicine, University of Pisa

Copyright

© BioMed Central Ltd 2001

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