According to the CDC, cancer deaths in the United States dropped 27% in the past 21 years, but still, it is the second leading cause of death. In traditional folk medicine, compounds extracted from various plant components such as fruit, seeds, leaves, flowers, etc., play a crucial role in treating several ailments, including cancer. Therefore, there is a constant interest in developing more effective, targeted, and cost-effective anticancer drugs. Medicinal plants are an enormously rich source of biologically active peptides and proteins. Nigella sativa, a miraculous medicinal herb commonly known as black seed or Kalonji, belongs to the plant family Ranunculaceae. Our group has previously reported the anticancer activity of proteins extracted from its seeds against the MCF7 breast cancer cell line. The present study focuses on applying two-dimensional liquid chromatography for protein purification and employing both LC/MS/MS and automated Edman sequencing techniques for the primary structure elucidation of purified proteins. The proteins were extracted in PBS, precipitated in 80% ammonium sulfate. The HiTrap Heparin column was employed to enriched the heparin-binding proteins/peptides from the crude extract and was were further purified by the second-dimensional RP-HPLC. The identity of several proteins was established by mass spectrometry and N-terminal amino acid sequencing. Our preliminary findings based on the homology search identified several biologically active proteins, including non-specific Lipid Transfer Protein (ns-LTP), Defensin, and Nigellin-1 proteins. Further, work is in progress on determining the complete primary structure of these and other biologically active proteins from the black seeds for their anticancer potentials in MCF-7 breast cancer and As-PC1 pancreatic cancer cell lines.

Purification and characterization of black seeds <i>(Nigella Sativa) </i>proteome

ACS Spring 2022

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Purification and characterization of black seeds (Nigella Sativa) proteome