Purification and characterization of non-enzymatic glycoprotein (NEGp) from flax seed buffer extract that exhibits anticoagulant and antiplatelet activity

Nandish, Sharath Kumar M. and Kengaiah, Jayanna and Ramachandraiah, Chethana and Chandramma and Shivaiah, Ashwini and Thirunavukkarasu and Shankar, Rohith L. and Sannaningaiah, Devaraja (2020) Purification and characterization of non-enzymatic glycoprotein (NEGp) from flax seed buffer extract that exhibits anticoagulant and antiplatelet activity. International Journal of Biological Macromolecules, 163. pp. 317-326. ISSN 1879-0003

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Abstract

The current study deals with the purification and characterization of non-enzymatic glycoprotein (NEGp) from flax seed buffer extract. Sephadex G-100 and DEAE-A25 column chromatography techniques were employed to isolate NEGp. NEGp showed single sharp band at 29 kDa region on 10% SDS-PAGE, and under reduced and non-reduced conditions revealed its monomeric nature. Besides, NEGp taken up the PAS stain at 29 kDa region reveals the presence of carbohydrate moiety. Purity of NEGp was adjudged by RP-HPLC, as it revealed a single sharp peak at the retention time of 3.4 min. The exact molecular mass of NEGp was found to be 26 kDa which was confirmed by MALDI-TOF. Circular di-chromism spectra of NEGp showed 12.0% alpha-helix, 24.3% alpha-helix turn and 63.7% random coils without beta pleated sheets. NEGp was found to exhibit anticoagulant activity by extending clotting time of both platelet rich plasma and platelet poor plasma from control 240 s to 1800 s and 280 s to 2100 s respectively at the concentration of 8 mu g. NEGp inhibited the agonists such as ADP, epinephrine and arachidonic acid induced platelet aggregation in washed platelets. The percentage of inhibition was found to be 70%, 80% and 60% respectively. While, it did not interfere in thrombin, PAF and collagen induced platelet aggregation. NEGp did not hydrolyse RBC membrane, devoid of haemorrhagic and edema inducing properties in experimental mice. (C) 2020 Elsevier B.V. All rights reserved.

Item Type: Article
Uncontrolled Keywords: Non-enzymatic glycoprotein; Anticoagulant; Antiplatelet and non-toxic
Subjects: C Chemical Science > Biochemistry
Divisions: Department of > Biochemistry
Depositing User: Mr Umendra uom
Date Deposited: 29 Mar 2021 06:19
Last Modified: 29 Mar 2021 06:19
URI: http://eprints.uni-mysore.ac.in/id/eprint/15474

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