Buffalo colostrum β-lactoglobulin inhibits VEGF-induced angiogenesis by interacting with G protein-coupled receptor kinase

Chougule, R. A. and Shilpa, P. and Bharathi P. Salimath and Aparna, H. S. (2013) Buffalo colostrum β-lactoglobulin inhibits VEGF-induced angiogenesis by interacting with G protein-coupled receptor kinase. Applied Biochemistry and Biotechnology, 171 (2). pp. 366-381. ISSN 1559-0291

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Abstract

β-lactoglobulin (β-lg), a major whey protein was purified and characterised from buffalo colostrum. The in silico analysis of the tryptic peptides based on LC-CID-MS/MS facilitated the identification of protein as β-lg. The sequences IIVTQ f1-5 and LSFNPTQLEEQCHV f(149-162) of m/z 933+ and 8512+ were found to match N- and C-extreme of β-lg while IDALNENK f(84-91) and TPEVDDEALEKFDK f(125-138) sequences deduced for m/z 916+ and 8182+ were in compliance to buffalo milk β-lg. Considering the sequence similarity of β-lg to glycodelin, a proven angiogenic protein, similar role for β-lg from buffalo colostrum (BLG-col) was examined. Interestingly, BLG-col exhibited anti-angiogenic activity by potently inhibiting cell proliferation, micro-vessel sprouting, cell migration and tube formation of human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner but having varied effect on Ehrlich ascites tumor cells, MCF-7, MDA-MB 435 and MDA-MB 231 cell lines. The anti-angiogenic potential of BLG-col was found to be vascular endothelial growth factor mediated. The immunolocalisation of BLG-col on the cell surface of HUVECs evidenced using FITC-labelled β-lg antibody indicated its extra-cellular binding. Furthermore, BLG-col interacting HUVEC membrane protein (64 kDa) was detected by immunoblot and its identity was established by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry analysis, which showed peptide sequence homology to G protein-coupled receptor kinase 4.

Item Type:	Article
Uncontrolled Keywords:	article, male, animal experiment, animal model, controlled study, nonhuman, rat, Animals, Rats, in vitro study, Humans, Male, gel permeation chromatography, sequence analysis, Cell Line, Cell Membrane, cell migration, Cell Movement, cell proliferation, Cell Proliferation, microvasculature, Neovascularization, Peptides, protein protein interaction, Tumor, Vascular Endothelial Growth Factor A, antiangiogenic activity, Protein Binding, vasculotropin, Physiologic, flow rate, sprouting, antiproliferative activity, Amino Acid Sequence, buffalo, Buffaloes, colostrum, Colostrum, computer model, Lactoglobulins, Ehrlich ascites tumor cell, Cell membranes, Mass spectrometry, Enzymes, amino acid sequence, Cell culture, angiogenesis, immunoblotting, endothelium cell, Human Umbilical Vein Endothelial Cells, protein localization, cell strain MCF 7, membrane protein, angiogenic protein, Anti-angiogenic activity, binding kinetics, Buffalo colostrum, Cell engineering, Cell proliferation, cell surface, competitive inhibition, Endothelial cells, G protein coupled receptor kinase, G protein coupled receptor kinase 4, G protein-coupled receptor kinase, G-Protein-Coupled Receptor Kinase 4, GRK 4, Human umbilical vein endothelial cells, immunocytochemistry, lactoglobulin, Lactoglobulin, Matrix assisted laser desorption, placenta protein 14, Proteomics, time of flight mass spectrometry, umbilical vein endothelial cell, Vascular endothelial growth factor, whey
Subjects:	B Life Science > Biotechnology
Divisions:	Department of > Biotechnology
Depositing User:	Arshiya Kousar Library Assistant
Date Deposited:	21 Sep 2019 09:49
Last Modified:	21 Dec 2020 05:16
URI:	http://eprints.uni-mysore.ac.in/id/eprint/7973

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