Preliminary characterization of a structural defect in homozygous sickled cell alpha spectrin demonstrated by a rabbit autoantibody

Carlos A. Monteiro, Xunda Gibson, Archil Shartava, Steven Goodman

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)200-205
Number of pages6
JournalAmerican Journal of Hematology
Volume58
Issue number3
DOIs
StatePublished - Jul 1 1998

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Spectrin
Autoantibodies
Rabbits
Erythrocytes
Dithiothreitol
Skeleton
Polyacrylamide Gel Electrophoresis
Membrane Proteins
Cell Count
Sickle Cell Trait
Reducing Agents
Indirect Fluorescent Antibody Technique
Actins
Molecular Weight
Western Blotting
Staining and Labeling

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Preliminary characterization of a structural defect in homozygous sickled cell alpha spectrin demonstrated by a rabbit autoantibody. / Monteiro, Carlos A.; Gibson, Xunda; Shartava, Archil; Goodman, Steven.

In: American Journal of Hematology, Vol. 58, No. 3, 01.07.1998, p. 200-205.

Research output: Contribution to journalArticle

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