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Buffalo Colostrum β-lactoglobulin Inhibits VEGF-Induced Angiogenesis by Interacting with G protein-Coupled Receptor Kinase

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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 f[1–5] 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.

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Acknowledgements

We thank University Grant Commission, New Delhi, India for sanction of grant (F31-294/2005-06) to undertake this investigation and Rohit A. Chougule thank ICMR for award of SRF.

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  1. Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore, 570 006, India

    Rohit A. Chougule, Shilpa P., Bharathi P. Salimath & Aparna H. Sosalegowda

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  1. Rohit A. Chougule
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Correspondence to Aparna H. Sosalegowda.

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Chougule, R.A., P., S., Salimath, B.P. et al. Buffalo Colostrum β-lactoglobulin Inhibits VEGF-Induced Angiogenesis by Interacting with G protein-Coupled Receptor Kinase. Appl Biochem Biotechnol 171, 366–381 (2013). https://doi.org/10.1007/s12010-013-0344-6

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