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Plasma anti-α-galactoside antibody binds to serine- and threonine-rich peptide sequence of apo(a) subunit in Lp(a)

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Abstract

Lipoprotein(a) immune complexes [Lp(a) IC] of varying particle density obtained by ultracentrifugation of plasma from normal healthy donors were markedly dominated by IgG. Lp(a) and immunoglobulins were liberated from plasma Lp(a) IC by treatment with melibiose, a sugar specific for circulating anti-α-galactoside antibody (anti-Gal). Upon incubation with plasma lipoprotein fraction anti-Gal but not the α-glucoside-specific antibody from human plasma formed de novo IC with Lp(a). Binding of Lp(a) sugar-reversibly enhanced the fluorescence of FITC-labeled anti-Gal as did binding of α-galactoside-containing glycoproteins. This effect apparently due to conformational shift in the Fc region of the antibody was also produced by apo(a) subunit separated from Lp(a) and de-O–glycosylated apo(a) but not by any other plasma lipoproteins or by Lp(a) pre-incubated with the O–glycan-specific lectin jacalin. O–Glycans and their terminal sialic acid moieties in apo(a) of circulating Lp(a)-anti-Gal IC, in contrast to those in pure Lp(a), were inaccessible to jacalin and anion exchange resin, respectively. Unlike other plasma lipoproteins, Lp(a) inhibited Griffonia simplicifolia isolectin B4 which also accommodates serine- and threonine-rich peptide sequence (STPS) as surrogate ligand to α-galactosides at its binding site. Results suggest that anti-Gal recognizes STPS in the O-glycan-rich regions of apo(a) subunit in Lp(a) which contains no α-linked galactose.

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Abbreviations

TAG:

Terminal α-linked galactose

STPS:

Serine- and threonine-rich peptide sequence

APAG:

Affinity-purified anti-Gal

GS I-B4 :

Griffonia simplicifolia isolectin B4

TI:

Soybean trypsin inhibitor

TIM:

Soybean trypsin inhibitor-melibiose

DIg:

Dextran-binding immunoglobulin

PBS:

Potassium phosphate buffer (20 mM) containing 150 mM NaCl

PBS-T:

PBS containing 0.05 % Tween 20

IC:

Immune complex

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Acknowledgments

Authors are grateful to Dr. Jaisy Mathai, Head, Department of Blood Transfusion Services of this institute for the out-dated plasma samples provided.

Conflict of interest

There is no conflict of interest of any sort in respect of any author or host institution.

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Authors and Affiliations

  1. Department of Biochemistry, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, 695011, India

    M. Geetha, V. Kalaivani, P. S. Sabarinath & P. S. Appukuttan

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  1. M. Geetha
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  2. V. Kalaivani
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  3. P. S. Sabarinath
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  4. P. S. Appukuttan
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Correspondence to P. S. Appukuttan.

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Geetha, M., Kalaivani, V., Sabarinath, P.S. et al. Plasma anti-α-galactoside antibody binds to serine- and threonine-rich peptide sequence of apo(a) subunit in Lp(a). Glycoconj J 31, 289–298 (2014). https://doi.org/10.1007/s10719-014-9521-2

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