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Spectrin interactions with globin chains in the presence of phosphate metabolites and hydrogen peroxide: implications for thalassaemia

Journal of Biosciences volume 32, pages 1147–1151 (2007)Cite this article

Abstract

We have shown the differential interactions of the erythroid skeletal protein spectrin with the globin subunits of adult haemoglobin (HbA); these indicate a preference for α-globin over that for β-globin and intact HbA in an adenosine 5′-triphosphate (ATP)-dependent manner. The presence of Mg/ATP led to an appreciable decrease in the binding affinity of the α-globin chain to spectrin and the overall yield of globin-spectrin cross-linked complexes formed in the presence of hydrogen peroxide. Similar effects were also seen in the presence of 2-,3-diphosphoglycerate (2,3 DPG), the other important phosphate metabolite of erythrocytes. The binding affinity and yield of cross-linked high molecular weight complexes (HMWCs) formed under oxidative conditions were significantly higher in α-globin compared with intact haemoglobin, HbA and the β-globin chain. The results of this study indicate a possible correlation of the preferential spectrin binding of the α-globin chain over that of the β-globin in the haemoglobin disorder β-thalassaemia.

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Abbreviations

2,3-DPG:

2-,3-diphosphoglycerate

ATP:

adenosine 5′-triphosphate

CM:

carboxymethylchloride

DEAE:

diethylaminoethyl cellulose

DTT:

dithiothreitol

F-spectrin:

fluorescein-conjugated spectrin

HbA:

adult haemoglobin

HMWC:

high molecular weight complex

Kd :

binding dissociation constant

Mg/ATP:

ATP in presence of 10-fold molar excess of MgCl2

PMB:

p-hydroxymercuribenzoic acid sodium salt

PMSF:

phenylmethylsulphonyl fluoride

RD:

relative pixel density

SEM:

standard error of the mean

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Affiliations

  1. Biophysics Division, Saha Institute of Nuclear Physics, 1/AF, Bidhannagar, Kolkata, 700 064, India

    Poppy Datta & Abhijit Chakrabarti

  2. Strcutural Genomics Section, Saha Institute of Nuclear Physics, 1/AF, Bidhannagar, Kolkata, 700 064, India

    Abhijit Chakrabarti

  3. Thalassemia Foundation, Kolkata, 700 064, India

    Sudipa Chakrabarty & Amit Chakrabarty

Authors
  1. Poppy Datta
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  2. Sudipa Chakrabarty
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  3. Amit Chakrabarty
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  4. Abhijit Chakrabarti
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Correspondence to Abhijit Chakrabarti.

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Datta, P., Chakrabarty, S., Chakrabarty, A. et al. Spectrin interactions with globin chains in the presence of phosphate metabolites and hydrogen peroxide: implications for thalassaemia. J. Biosci. 32, 1147–1151 (2007). https://doi.org/10.1007/s12038-007-0116-y

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  • DOI: https://doi.org/10.1007/s12038-007-0116-y

Keywords

  • Globin subunits
  • erythroid spectrin
  • oxidative cross-linking
  • β-thalassaemia