Premium
Preliminary characterization of a structural defect in homozygous sickled cell alpha spectrin demonstrated by a rabbit autoantibody
Author(s) -
Monteiro Carlos A.,
Gibson Xunda,
Shartava Archil,
Goodman Steven R.
Publication year - 1998
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/(sici)1096-8652(199807)58:3<200::aid-ajh7>3.0.co;2-v
Subject(s) - spectrin , autoantibody , epb41 , dithiothreitol , red blood cell , tetramer , chemistry , microbiology and biotechnology , alpha (finance) , antibody , red cell , cell , biochemistry , cytoskeleton , biology , immunology , medicine , enzyme , construct validity , nursing , patient satisfaction
We have identified a rabbit autoantibody that strongly reacts with the core membrane skeleton of control red blood cells, and does not react with low‐ or high‐density sickle cell core skeletons upon indirect immunofluorescence. Western blot analysis of red blood cell membrane proteins, utilizing this autoantibody, indicated no reactivity to any protein when SDS‐PAGE was conducted in the presence of the reducing agent, dithiothreitol. However when SDS‐PAGE was performed on control red blood cell membrane proteins separated in the absence of dithiothreitol, the autoantibody specifically reacted with a high molecular weight polypeptide (apparent M r ≊ 310 kD) representing a DTT sensitive form of control α spectrin, which we refer to as α / spectrin. There was no staining of high density or low density sickle cell α or α / spectrin. This autoantibody should be an excellent tool for the fine mapping of structural change(s) in control vs. sickle cell α spectrin, and determination of whether the structural alteration effects spectrin dimer‐tetramer interconversion and/or the spectrin‐actin interaction. The modification in α spectrin, detected by this antibody, is very specific for homozygous SS α spectrin because sickle cell β + thalassemic α spectrin and sickle cell trait α spectrin react intensely with the autoantibody. Am. J. Hematol. 58:200–205, 1998. © 1998 Wiley‐Liss, Inc.