Nephrology Dialysis Transplantation, Vol 12, Issue 6 1192-1198, Copyright © 1997 by Oxford University Press
M Garcia-Garcia, A Gouin-Charnet, G Mourad and A Argiles
Background. There is a controversy as to whether
{beta}2-microglobulin ({beta}2M amyloid deposits may be degraded
resulting in regression and cure of amyloidosis. We have recently reported
a long-term clinical study involving transplanted patients suggesting that
there is no resorption of amyloid deposits in vivo,
even after correction of the primary cause of amyloidosis. To progress in
the study of the solubility of amyloid fibrils we performed an in
vitro study with the intent to remove protein constituents from
amyloid fibrils and amyloid deposits. Methods. Amyloid
fibrils were prepurified from four amyloid deposits surgically obtained
from carpal tunnel. They were incubated for 2 h with a phosphate-buffered
saline (PBS) solution containing trypsin, collagenase, kallikrein, the
three of them, or PBS alone. The experiments were repeated in the presence
of the antiprotease &agr;2M). Results. Several
bands were observed when the supernatants were run through SDS-PAGE.
Western blotting identified in these bands the presence of &agr;2M,
light chains of immunoglobulins and {beta}2M in mono- and dimeric form.
The same proteins were solubilized with PBS alone. Equivalent results were
obtained with crude amyloid deposits; however, {beta}2M presented almost
exclusively in monomeric form. Conclusions. These
results show that the protein constituents may be recovered from the
amyloid fibrils in vitro. They also show that even the
more insoluble {beta}2M dimers are resuspended by the action of PBS,
with no need for proteases to cleave their attachment to the amyloid
deposits. Keywords: amyloid resorption; proteases;
&agr;2-microglobulin; light chains of immunoglobulins amyloid
resorption; proteases; &agr;2-macroglobulin; {beta}2-microglobulin;
light chains of immunoglobulins
ORIGINAL ARTICLES
Monomeric and dimeric {beta}2-microglobulin may be extracted from amyloid deposits in vitro
Centre de Recherche Biochimie Macromoleculaire (CRBM), CNRS LP 9008, INSERM Unit 249, University of Montpellier I, BP 5051, route de Mende, 34033 Montpellier Cedex, France; Department of Nephrology, University Hospital Lapeyronie, Montpellier Cedex, France; Corresponding author
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  D.-P. Hong, M. Gozu, K. Hasegawa, H. Naiki, and Y. Goto Conformation of beta 2-Microglobulin Amyloid Fibrils Analyzed by Reduction of the Disulfide Bond J. Biol. Chem., June 7, 2002; 277(24): 21554 - 21560. [Abstract] [Full Text] [PDF]
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