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

Datta, Poppy; Chakrabarty, Sudipa Basu; Chakrabarty, Amit; Chakrabarti, Abhijit

doi:10.1007/s12038-007-0116-y

Cited by 11 publications

(20 citation statements)

References 23 publications

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“…The HS patients we have studied, perhaps carry dominant mutations in ankyrin, β-spectrin and band 3, drastically affecting PS asymmetry [24]. We have earlier reported higher spectrin binding affinity of α-globin chain compared to β-globin and enhanced yield of spectrin-globin-hydrogen peroxide cross-linked complexes in thalassemia [27,28]. Earlier work from our laboratory also indicated loss of red cell PS asymmetry in different cases of hematological malignancies [29].…”

Section: Discussionmentioning

confidence: 79%

Eryptosis in hereditary spherocytosis and thalassemia: role of glycoconjugates

Basu

Banerjee

Chandra³

et al. 2009

Glycoconj J

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The present work is aimed to study the mechanism of faster erythrocyte clearance in hereditary spherocytosis (HS), a heterogeneous disorders characterized by alterations in the proteins of the red cell membrane skeleton along with different kinds of thalassemia. The maximum exposure of phosphatidylserine (PS) is found in HS compared to those in both α- and β-thalassemia. Interestingly, in HS more PS exposed cells were found in younger erythrocytes compared to normal and the thalassemics where aged cells showed higher loss of PS asymmetry. Loss of sialic acid and GlcNAc bearing glycoconjugates, presumably the glycophorins, was also found upon aging. The loss of PS asymmetry together with the cell surface glycoproteins mediated by membrane vesiculation, seemed to play key role in early clearance of erythrocytes from circulation following a mechanism similar to HbEβ-thalassemia.

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Section: Discussionmentioning

confidence: 79%

Eryptosis in hereditary spherocytosis and thalassemia: role of glycoconjugates

Basu

Banerjee

Chandra³

et al. 2009

Glycoconj J

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“…Globin chain imbalance could be much more severe in the younger erythrocytes of HbEβ‐thalassaemia and also a probable cause of higher PS exposure. We have previously reported the higher spectrin‐binding affinity of HbE and the α‐globin chain when compared with HbA and β‐globin respectively, and enhanced yield of spectrin‐globin‐hydrogen peroxide cross‐linked complexes in HbEβ‐thalassaemia (Datta et al , 2006, 2007). Taken together, all of these could contribute to the loss of PS asymmetry in HbEβ‐thalassaemic erythrocytes.…”

Section: Discussionmentioning

confidence: 99%

Loss of phospholipid membrane asymmetry and sialylated glycoconjugates from erythrocyte surface in haemoglobin E β‐thalassaemia

Basu

Banerjee

Chandra³

et al. 2008

Br J Haematol

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Summary This study aimed to investigate any correlation between the extent of phosphatidylserine (PS) asymmetry and sialylated glycoconjugate levels with the faster clearance of circulating erythrocytes in haemoglobin E (HbE) β‐thalassaemia. Erythrocytes from peripheral blood samples of different HbEβ‐thalassaemia patients showed loss of PS asymmetry measured by annexin V binding using flow cytometry. Maximum PS exposure was found when HbE was 50–60% and HbF was <20% indicating a possible correlation with severity of the disease. Separation of erythrocytes into aged and younger cells showed higher loss of PS asymmetry in the younger erythrocytes of HbEβ‐thalassaemia patients when compared with normal blood, where PS asymmetry was lost only in the older cells. Sialylated glycoconjugate measurement using the lectins wheatgerm agglutinin and pokeweed mitogen showed loss of sialic acid and N‐acetyl‐d‐glucosamine‐bearing glycoproteins in the order normal

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“…Presence of oxy form of hemoglobin was confirmed using absorbance at 415 nm and 541 nm. Hemoglobin preparation was stored at −70 °C for a maximum of 1 week (Datta, Chakrabarty, Chakrabarty, & Chakrabarti, ).…”

Section: Methodsmentioning

confidence: 99%

“…α and β globin subunits were isolated from hemoglobin using previously published protocol (Datta et al, ; Datta et al, ). Briefly, 100 mg PMB per 1 g of hemoglobin was dissolved in minimum volume of 0.1 M KOH and 1 M acetic acid was added till very light precipitate persisted.…”

Section: Methodsmentioning

confidence: 99%

“…Dimeric human erythroid spectrin was isolated from clean white RBC ghost membranes following protocol elaborated in earlier studies (Datta, Chakrabarty, Chakrabarty, & Chakrabarti, 2003). Briefly Hemoglobin preparation was stored at −70 C for a maximum of 1 week (Datta, Chakrabarty, Chakrabarty, & Chakrabarti, 2007).…”

Section: Isolation Of Human Erythroid Spectrinmentioning

confidence: 99%

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Localizing the chaperone activity of erythroid spectrin

Bose

Chakrabarti

2019

Cytoskeleton

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Spectrin, the major protein of the erythrocyte membrane skeleton has canonically been thought to only serve a structural function. We have previously described a novel chaperone‐like property of spectrin and also hypothesized that the chaperone activity and binding of a hydrophobic ligand, Prodan are localized in the self‐association domain. Here we probe the location and molecular origin of the chaperone activity of multi‐domain spectrin using a selection of individual recombinant spectrin domains, which we have characterized using intrinsic tryptophan fluorescence and CD spectroscopy to show their identity to native spectrin. Aggregation assays using insulin, ADH, α‐ and β‐globin as well as enzyme refolding assays using alkaline phosphatase and α‐glucosidase show that the chaperone activity is not only localized in the self‐association domain but is a generalized property of spectrin domains. This is to our understanding, a unique feature in the case of modular multi‐repeat proteins, possibly implicating that the large family of “spectrin‐repeat” domain containing proteins may also have chaperone like property. Substrate selectivity of chaperone activity as evidenced by the preferential protection of α‐ over β‐globin chains is seen; which has implications in hemoglobin diseases. Moreover, enzyme‐refolding assays also indicate alternate modes of chaperone action. We propose that the molecular origin of chaperone activity resides in the surface exposed hydrophobic patches of the spectrin domains as shown by ANS (1‐anilinonaphthalene‐8‐sulfonic acid) and Prodan (6‐propionyl‐2[dimethylamino]‐naphthalene) binding. We also show that Prodan does indeed have a unique binding site on spectrin located at the self‐association domain.

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

Cited by 11 publications

References 23 publications

Eryptosis in hereditary spherocytosis and thalassemia: role of glycoconjugates

Eryptosis in hereditary spherocytosis and thalassemia: role of glycoconjugates

Loss of phospholipid membrane asymmetry and sialylated glycoconjugates from erythrocyte surface in haemoglobin E β‐thalassaemia

Localizing the chaperone activity of erythroid spectrin

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