IDENTIFICATION OF A REGION OF BETA-2-GLYCOPROTEIN-I CRITICAL FOR LIPID-BINDING AND ANTICARDIOLIPIN ANTIBODY COFACTOR ACTIVITY

HUNT, JE; Simpson, RJ; KRILIS, SA

Author(s): HUNT, JE; Simpson, RJ; KRILIS, SA;
Details: Publication Year 1993-03-15,Volume 90,Issue #6,Page 2141-2145
Journal Title: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Publication Type: Journal Article
Abstract: Beta2-glycoprotein I (beta2-GPI), a phospholipid-binding plasma protein, is an absolute requirement (cofactor) for the binding of autoimmune-type anti-cardiolipin (aCL) antibodies to cardiolipin (CL). The nature of this cofactor activity and the specific regions of the molecule involved have not yet been determined. We have identified a preparation of beta2-GPI that lacks aCL antibody cofactor activity. Analysis of the structural differences between the active and inactive forms enabled identification of the region of beta2-GPI critically important for aCL cofactor activity. The active form of beta2-GPI bound CL and displayed cofactor activity down to 1 mug/ml. The inactive form failed to bind CL and possessed no cofactor activity even at concentrations up to 94 mug/ml, indicating that the ability of beta2-GPI to bind lipid is an absolute requirement for aCL cofactor activity. Both forms possessed identical N-terminal sequences and were recognized as essentially immunoreactively identical by polyclonal antisera to beta2-GPI. However, the inactive form has undergone proteolytic cleavage and exists primarily as a ''clipped'' molecule, the polypeptide chain being cleaved between Lys-317 and Thr-318 (a potential thrombin cleavage site), with the two cleaved segments linked as a disulfide-bonded complex. This indicates that the C-terminal region is critically important for beta2-GPI to bind lipid and for aCL cofactor activity. The clipped form of beta2-GPI would not be suitable for use as aCL cofactor and its use may have led some investigators to conclude incorrectly that beta2-GPI does not interact with aCL antibodies.
Publisher: NATL ACAD SCIENCES
Keywords: SYSTEMIC LUPUS-ERYTHEMATOSUS; AMINO-ACID-SEQUENCE; ANTIPHOSPHOLIPID ANTIBODIES; APOLIPOPROTEIN-H; HUMAN-SERUM; PLATELETS; ANTICARDIOLIPIN; PURIFICATION; CLEAVAGE
Publisher's Version: https://doi.org/10.1073/pnas.90.6.2141
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 1993-03-15 12:00:00