Nova Biotechnol Chim (2020) 19(1): 37-51 DOI 10.36547/nbc.v19i1.576  Corresponding author: toluwase.fatoki@fuoye.edu.ng, fheztbioinformatics@gmail.com Nova Biotechnologica et Chimica In silico study of anticarcinogenic potential of the selenoprotein BthD from Drosophila melanogaster. Identifying the anticancer peptide CRSUR from the conserved region Toluwase Hezekiah Fatoki1,2,, Omodele Ibraheem1, Amos Olalekan Abolaji3 and David Morakinyo Sanni4 1Translational Bioinformatics Unit, Department of Biochemistry, Federal University Oye, PMB 373 Oye-Ekiti, Ekiti State, Nigeria 2Fhezt Bioinformatics Laboratory, Ifaki-Ekiti, Ekiti State, Nigeria 3Department of Biochemistry, Faculty of Basic Medical Sciences, College of Medicine, University of Ibadan, Oyo State, Nigeria 4Department of Biochemistry, Federal University of Technology Akure, PMB 704, Akure, Ondo State, Nigeria Article info Article history: Received: 28th February 2020 Accepted: 5th May 2020 Keywords: Anticancer peptide Drosophila melanogaster Mechanism design NF-kappa-B signaling Selenoprotein BthD Abstract Drosophila melanogaster is used as a model system in biomedical studies. Selenoprotein is the major biological form of selenium in eukaryotes. Selenoproteins are generally involved in catabolic pathways in bacteria and archaea, whereas it participates in anabolic and antioxidant processes in eukaryotic. In this study, anticancer potential of selenoprotein BthD of D. melanogaster was investigated using bioinformatics methods. Results showed that selenoprotein BthD of D. melanogaster may have dual properties as evident by its orthology with selenoprotein H (SelH) of Homo sapiens and conserved domain of fructokinase-like protein 2 of Vitis vinifera. These dual properties were also revealed in the phylogenetic analysis, while further structural modeling showed that selenoprotein BthD possibly exists as homotetramer in the native functional structure. The anticancer property of selenoprotein BthD was proposed to be by synergy of antioxidant or redox activities of thioredoxin and glutathione reductase (TGR) domain and the signaling function of fructokinase-like protein 2 domain both in Golgi apparatus and cytoplasm, through energy deprivation. The anticancer peptide CRSUR was identified from conserved region of selenoprotein BthD, of which its cyclic form showed potential anticancer properties predictively through E3 ubiquitin-protein ligase regulating NF-kappa-B signaling by unleashing cells for spontaneous formation of the ripoptosome.  University of SS. Cyril and Methodius in Trnava Introduction The genomic sequence of D. Melanogaster is about 115229998 bp and contains 13329 annotated genes (Adams et al. 2000). Drosophila possess genes systems which regulate nutrient uptake, storage and metabolism that are critical to survival and have been found conserved almost in all eukaryotes including humans, brown alga, zebrafish, mouse, Escherichia coli, and Caenorhabditis elegans (Allocca et al. 2018; Hatfield et al. 2014). D. melanogaster has been used as a model system for toxicological studies and diseases mechanism such as neurological mailto:toluwase.fatoki@fuoye.edu.ng mailto:fheztbioinformatics@gmail.com Nova Biotechnol Chim (2020) 19(1): 37-51 38 disorders, developmental disorders, metabolic and storage disorders, cancer and cardiovascular disease (Bier 2005; Abolaji et al. 2014; Saraiva et al. 2018). The major biological form of selenium in eukaryotes is selenocysteine (Sec) and its mostly found in the active site of selenoproteins. Selenocysteine is called the 21st amino acid which has chemical structure differs from cysteine only by the presence of selenium in place of the sulfur atom. Sec is co-translationally inserted into a polypeptide chain in response to in-frame UGA codons directed by the Sec insertion sequence element, a stem-loop structure in the untranslated regions (3-UTRs) of selenoprotein mRNAs (Hatfield et al. 2014). The human genome contains 25 selenoprotein genes (Kryukov et al. 2003) and they are involved in a variety of functions, most notably redox homeostasis. Larger selenoproteomes can be found in aquatic organisms such as zebrafish, and brown alga (Lobanov et al. 2007; Hatfield et al. 2014). Selenoproteins are generally involved in catabolic pathways in bacteria and archaea, whereas eukaryotic selenoproteins participate rather in anabolic and antioxidant processes (Herbette et al. 2007). Selenoproteins participate in thyroid hormone metabolism, muscle formation, selenocysteine synthesis and in sperm maturation (Rederstorff et al. 2006). According to Gladyshev et al. (2016), selenoproteins without known functions include SELENOF (selenoprotein F, 15-kDa selenoprotein, SEP15), SELENOH (selenoprotein H, SELH, C11orf31), SELENOK (selenoprotein K, SELK), SELENOM (selenoprotein M, SELM), SELENOO (selenoprotein O, SELO), SELENOT (selenoprotein T, SELT), and others. Human selenoprotein enzymes with known functions such as thioredoxin reductases (TR1), glutathione peroxidases (Sep15 and GPx2) are important cellular redox-regulators needed by both normal and cancer cells, which result in anti- and pro- tumorigenic effects at a tissue-specific cellular level (Hatfield et al. 2014). In liver TRI exhibits anticancer protein and in lung TR1 is a pro-cancer protein and a prime candidate for cancer therapy (Yoo et al. 2006; Carlson et al. 2012). The GPx1 polymorphisms are associated with cancer risk (Zhuo and Diamond 2009). It remains to be elucidated whether these anti- and pro-tumorigenic effects are tumor stage or grade-dependent. The known selenoproteins in D. melanogaster are dSPS2, dSelK (former called dSelG) and dSelH (also known as dSelM or BthD) (Hirosawa- Takamori et al. 2000; Castellano et al. 2001). dSelK has one cysteine homolog and dSelH has two (Castellano et al. 2001; Martin-Romero et al. 2001). dSelH appears to belong to a class of selenoproteins widely distributed across the phylogenetic spectrum, as dSelH was found in zebrafish, human and mouse expressed sequence tag (EST) databases (Dickiy et al. 2007; Novoselov et al. 2007). The ability of dselH to reverse the toxic effects of glutathione depletion in Schneider cells was suggested to reflect a glutathione sparing effect via increased activity of an alternative anti-oxidant pathway, which restores the perturbed anti-oxidant-pro-oxidant balance (Morozova et al. 2003). Disruption of selenophosphate synthetase expression has recently been shown to modulate the Ras/MAPK signalling cascade in flies (Morey et al. 2001). Ser/Thr kinase domain is one of the core kinase cascades in D. melanogaster and mammals (Yin and Zhang 2011). In D. melanogaster, Ser/Thr kinase domain is found in the core kinases of Hippo signaling pathway, as well as in the four- jointed and discs overgrown of upstream regulatory components (Yin and Zhang 2011). Defects in the core kinases and some of the upstream components of the pathway lead to robust organ overgrowth and link to numerous cancers (Pan 2010; Zhang et al. 2009). The Hippo signaling pathway limits organ size by regulating cell proliferation and apoptosis. In this study, anticancer potential of selenoprotein BthD of D. melanogaster was investigated based on the available genomic data and publications. The gene product with anticancer properties in an insect could be useful in the development of biologic agent for human cancer therapy. Experimental Drosophila melanogaster selenoproteins The selenoprotein was searched from D. melanogaster database on Ensembl genome browser v97 (http://www.ensembl.org). The genes http://www.ensembl.org/ Nova Biotechnol Chim (2020) 19(1): 37-51 39 obtained from ensembl were looked up in the D. melanogaster database (www.flybase.org). The protein sequences and information were obtained from UniProt database (www.uniprot.org). The protein sequence of BthD of D. melanogaster was used to query Homo sapiens database on the Blastp server of NCBI (Camacho et al. 2009). Conserved protein domain Human selenoproteins TRI, Sep15 and GPx2 has been reported to possessed anti- and pro- tumorigenic effect (Hatfield et al. 2014). The sequence of these proteins was obtained from UniProt. The conserved protein domain of all the selenoproteins of D. Melanogaster and four selenoproteins of Homo sapiens were investigated using the protein sequences on the CDD server (https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb .cgi) of NCBI (Marchler-Bauer et al. 2017). The protein sequence of BthD of D. melanogaster was used to query database of eight plants in the taxonomy of Mesangiospermae identified from CDD results, on the Blastp server of NCBI (Camacho et al. 2009). Phylogenetics analysis The protein sequence of selenoproteins of D. melanogaster and H. sapiens obtained from the previous steps, were used for phylogenetics study. Multiple sequence alignment on ClustalO server (www.ebi.ac.uk/tools/msa/clustalo) was carried out and phylogenetic tree was constructed. The tree data was visualized at www.phylo.io. Structural modeling of selenoprotein BthD The three-dimensional structure of selenoprotein BthD of D. melanogaster was modelled on Swissmodel server (www.swissmodel.ch) using protein sequence (Camacho et al. 2009; Remmert et al. 2012; Waterhouse et al. 2018). Integration of anticarcinogenic mechanism of selenoprotein BthD The D. melanogaster pathways associated with growth of tumor were obtained from www.kegg.jp and available information in the scientific literatures was mined. These data were integrated with the key results of this study and anticarcinogenic mechanism of selenoprotein BthD of D. melanogaster was proposed. In silico prediction anticancer peptides of selenoprotein BthD and their physicochemical properties The prediction of anticancer peptides in selenoprotein BthD was performed on AntiCP server Protein Scan (https://webs.iiitd.edu.in/raghava/anticp/submit_pro t.php), and generated the fragments amino acids residues of length of 5 with minimum Support vector machine (SVM) score of 1.15, and predict their anticancer property along with all Table 1. Details of selenoprotein genes, and proteins of D. melanogaster integrated from Ensembl, Flybase and Uniprot. Gene name Chromosome location No. of transcript Human orthologs [species\gene symbol] Best transcript name and ID Length [nt] Uniprot ID Length [amino acids] Subcellular location BthD Chr. X: 13,612,131- 13,613,228 1 Hsap\SELENOH BthD-RA, FBtr0073806 977 Q9VYB0 249 Cytoplasm SelG Chr. X: 11,887,284- 11,888,24 1 Hsap\SELENOK SelG-RA, FBtr0073570 827 Q7Z2C4 110 Golgi apparatus SelR Chr. 3R: 10,863,355- 10,868,642 8 Hsap\MSRB3 SelR-RE, FBtr0082295 1156 Q8INK9, D3DMP0 208, 192 Nucleus, Cytoplasm, cyto- skeleton SelT Chr. 2L: 5,010,922- 5,12,127 3 Hsap\SELENOT SelT-RA, FBtr0079000 960 M9PCL4, Q9VMV6 198 Integral component of membrane, endomembrane system http://www.flybase.org/ http://www.uniprot.org/ https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi http://www.ebi.ac.uk/tools/msa/clustalo http://www.phylo.io/ http://www.swissmodel.ch/ http://www.kegg.jp/ https://webs.iiitd.edu.in/raghava/anticp/submit_prot.php https://webs.iiitd.edu.in/raghava/anticp/submit_prot.php Nova Biotechnol Chim (2020) 19(1): 37-51 40 Fig. 1. Model structures of selenoprotein BthD of D. melanogaster in homotetramer oligomeric state. the important physico-chemical properties like hydrophobicity, charge, pI etc. The cell-penetrating efficacy of peptides were predicted on CPPred-RF server (http://server.malab.cn/CPPred-RF/index.jsp) (Wei et al. 2017). In silico target prediction and pharmacokinetics of anticancer peptides of selenoprotein BthD The structure and SMILES (Simplified Molecular Input Line Entry Specification) format of selenocysteine-containing peptide (in cysteine form) was obtained from PepSMI server (https://www.novoprolabs.com/tools/convert- peptide-to-smiles-string) and cysteine residue was edited to selenocysteine on https://pubchem.ncbi.nlm.nih.gov/edit3/index.html. In silico prediction of target was done on SwissTargetPrediction server, where Homo sapiens was selected as target organism (Diana et al. 2019). The active peptides (peptides with predicted target) were then subjected to in silico Absorption, Distribution, Metabolism, and Excretion (ADME) screening on SwissADME server at default parameters. (Diana et al. 2017). Results The search on the Ensembl genome browser showed four genes of selenoproteins present in the D. melanogaster genome. The gene products, orthology in H. sapiens and subcellular location were obtained from Flybase and UniProt databases, as shown in Table 1. Each of the four selenoproteins in D. melanogaster has one ortholog in human. The homology alignment of BthD protein sequence (Uniprot ID: Q9VYB0) by Blastp confirmed 41.38 % similarity with human SelH (C11orf31) selenoprotein (Uniprot ID: Q8IZQ5) which is located in the Golgi apparatus and cytoplasm. The conserved domain of D. melanogaster and H. sapiens are summarized in Table 2. The conserved domain of selenoprotein BthD was found to be exceptional with no similarity in H. sapiens unlike the other three. The conserved protein domain of BthD was classified as kinase which belong to PLN02967 superfamily of fructokinase-like protein 2 (EC 2.7.1.4), and belong to protein clusters conserved in taxonomy of Mesangiospermae in eukaryotic plant. This study is the first to discover and report the plant-like properties of BthD. Further homology alignment of the BthD protein sequence against eight plants in the PLN02967 superfamily of taxonomy Mesangiospermae, only showed 33.33 % similarity to an uncharacterized selenoprotein H (UniProt ID: D7SU28) of Vitis vinifera. Moreover, human SelH contained domain architecture which is similar to that SelT of D. melanogaster and has been noted as thioredoxin and glutathione reductase (TGR Domain). This domain is homodimeric, FAD- containing member of the pyridine nucleotide disulfide oxidoreductase family. Table 3 shows the summary of homology analysis of BthD protein M1 M2 M3 http://server.malab.cn/CPPred-RF/index.jsp https://www.novoprolabs.com/tools/convert-peptide-to-smiles-string https://www.novoprolabs.com/tools/convert-peptide-to-smiles-string https://pubchem.ncbi.nlm.nih.gov/edit3/index.html Nova Biotechnol Chim (2020) 19(1): 37-51 41 Nova Biotechnol Chim (2020) 19(1): 37-51 42 Nova Biotechnol Chim (2020) 19(1): 37-51 43 Fig. 2. Multiple sequence alignment of all selenoproteins of D. melanogaster, four selenoproteins of H. sapiens and a selenoprotein of V. vinifera. sequence of D. melanogaster against H. sapiens and V. vinifera integrated from Ensembl Browser v97, Blastp of NCBI and UniProt. The structural model parameter of BthD selenoprotein on Swissmodel server is shown in Table 4. The structure was modeled as homotetramer based on the percentage similarities to three different template proteins; 2ojl (crystal structure of Q7WAF1_BORPA from Bordetella parapertussis), 2obk (crystal structure of the putative Se binding protein from Pseudomonas fluorescens), and 2oka (crystal structure of Q9HYQ7_PSEAE from Pseudomonas aeruginosa) as shown in Fig. 1. This structure can be further elucidated through X-ray crystallography. The result in Fig. 2 showed the position of conserved amino acid residues among selenoproteins (D7SU28_ VITVI, BTHD_DROME and SELH_HUMAN). The phylogenetic tree confirmed the evolutionary relatedness of BthD gene of D. melanogaster, SelH gene of H. sapiens and D7SU28 gene of V. vinifera (Fig. 3). The integral mechanism of anticarcinogenic property of selenoprotein BthD is shown in Fig. 4. This mechanism was based on the glycolytic energy stress induced by hepatocytes depletion of ATP by fructose and its impact of AMPK1, Hippo signaling and Notch1 signaling pathways; the role of trehalose to regulate glucose metabolism in stress condition; the ability of isomaltose to regulate adenylate biosynthesis; and the capacity V. vinefera to provide fructose and phytochemicals such as resveratrol. Based on the mechanism proposed in this study, we have hypothesized that amino acid sequences EHCRSUR and GAPRRGA from selenoprotein BthD as well as EHCKQCN and EKPRRGC from V. vinefera (grape) could be the key bioactive peptides that can alone and synergistically modulate this mechanism and impact the required antioxidant effect. This was validated by the results of anticancer peptides from selenoprotein BthD shown in Table 5, where selenocysteine-containing peptide CRSUR has SVM score of 1.37 and high Nova Biotechnol Chim (2020) 19(1): 37-51 44 Fig. 3. The phylogenetic tree of all selenoproteins of D. melanogaster, four selenoproteins of H. sapiens and a selenoprotein of V. vinifera. Cell-penetrating uptake efficiency, while RRGAF has SVM score of 1.28 and high cell-penetrating uptake efficiency. The structure of the CRSUR peptide in linear and cyclic (head-to-tail bond) is shown in Table 6. The predicted targets of CRSUR peptide from selenoprotein BthD show that this selenocysteine-containing peptide have anticancer and antiviral or antineuronal properties based on the biological processes by the target genes obtained such as Furin, Integrin beta- 3, andBaculoviral IAP repeat-containing protein (Table 7). The result of pharmacokinetics of CRSUR (Table 8) indicate that gastrointestinal route will not be good for the administration of bioactive peptide. Discussion Selenoprotein BthD (BthD) has been reported to possesses antioxidant potential (Castellano et al. 2001). Selenoprotein T (SelT) is a thioredoxin- disulfide reductase (EC 1.8.1.9) that belongs to the SelWTH family and SELT subfamily. Selenoprotein R (SelR or MsrB) is a peptide- methionine (R)-S-oxide reductase (EC 1.8.4.12) which belongs to the MsrB Met sulfoxide reductase family (Kryukov et al. 2003). The thioredoxin and glutathione reductase (TGR Domain) is homodimeric, FAD-containing member of the pyridine nucleotide disulfide oxidoreductase family which contains a C-terminal motif Cys- SeCys-Gly, where SeCys is selenocysteine encoded by TGA which is a stop codon in some sequence). (Sun et al. 2001 TIGR02174 domain is a member of the superfamily cl01407 together with related to pfam10262, a domain found in both bacteria and animals selenoproteins SelT, SelW, and SelH (Dickiy et al. 2007). Taxonomy of Mesangiospermae belongs to eukaryotic plant and its PLN02967 superfamily was similar to an uncharacterized selenoprotein H (UniProt ID: D7SU28) of V. vinifera. Some studies have reported that grape (V. vinifera) extracts showed cytotoxicity towards cultured cells as well as inhibited tumor growth in animal models (Shrotriya et al. 2012; Sun et al. 2012). Different molecular mechanisms have been proposed for these protective effects of grape extracts, such as inhibition of enzymes playing an essential role in cell proliferation (e.g. human topoisomerase I) and inhibition of angiogenesis (Agarwal et al. 2004; Stagos et al. 2005). The result of a double-blinded randomized crossover human trial showed that dietary supplementation of grape seed extract at a dose of 600 mg/day for 4 weeks can decrease of oxidative stress and enhance glutathione (GSH)/oxidized glutathione (GSSG) and total antioxidant status (Kar et al. 2009). The anticancer effects of whole black grape (seeds included) extract have been reported in the cancerous colon tissues of humans by inhibition in DNA turnover enzymes (Durak et al. 2005). An in silico study has reported the molecular targets for the key bioactive components present in grape such as resveratrol, piceatannol, and scirpusin A (Fatoki et al. 2018a). Nova Biotechnol Chim (2020) 19(1): 37-51 45 Fig. 4. Integrated anticarcinogenic mechanism of Selenoprotein BthD. Comprehensive review of anticancer properties of grape can be found in another publication (Zhou and Raffoul 2012). In D. melanogaster, three forms of hexokinases (Hex A, B and C) were found by agar gel electrophoresis (Madhavan et al. 1972). Hex A and Hex B have been mapped on the same structural gene on the X chromosome (Voelker et al. 1978) while Hex C on the second chromosome (Jelnes 1971). However, whether these three hexokinases were either aldose or ketose was not investigated. The chromosomal location of BthD is Nova Biotechnol Chim (2020) 19(1): 37-51 46 Table 6. Structure of the peptide CRSUR Linear and Head-to-tail bond. Type Smiles Structure Linear N[C@@H](CS)C(=O)N[C@@H](CCCNC( =N)N)C(=O)N[C@@H](CO)C(=O)N[C@ @H](C[Se])C(=O)N[C@@H](CCCNC(=N) N)C(=O)O Head-to-tail bond N1[C@@H](CS)C(=O)N[C@@H](CCCNC (=N)N)C(=O)N[C@@H](CO)C(=O)N[C@ @H](C[Se])C(=O)N[C@@H](CCCNC(=N) N)C1=O Chromosome X: 13,612,131-13,613,228 (Table 1). Fructokinase (also known as ketohexokinase; KHK), which catalyzes the phosphorylation of fructose to fructose 1-phosphate, was identified by MALDI-TOF MS and found expressed at extremely low rates in the renal tumor tissues (Hwa et al. 2006). A study has shown that fructose- induced ATP depletion in human, rat and mouse hepatocytes cause full protected against tumor necrosis factor-alpha (TNF-α)-induced cytotoxicity, whereas hepatic tumor cell lines showed increased hexokinase II (HKII) expression which inhibited fructose-mediated cytoprotection (Speicher et al. 2010). Study has shown that trehalose do not stimulate rapid increases in blood glucose and excessive secretion of insulin and gastric inhibitory polypeptide (GIP) promoting fat accumulation (Yoshizane et al. 2017). It has been demonstrated that trehalose-6-phosphate (T6P) inhibits yeast hexokinase 2 (HKII) activity, thus it is likely that this metabolite regulates glycolysis by modulating the flow of phosphorylated sugars towards this pathway (Blázquez et al. 1993; Nova Biotechnol Chim (2020) 19(1): 37-51 47 Table 7. Predicted targets of CRSUR Peptide from selenoprotein BthD. S/N Targets UniProt ID CRSUR Peptide [% probability] Linear Head-to-tail bond 1 Furin P09958 60 45 2 Proprotein convertase subtilisin/kexin type 4, 5, 6 Q6UW60, Q92824, P29122 60 45 3 Complement factor B Ba fragment P00751 55 - 4 Complement C2 P06681 55 - 5 Neurotensin receptor type 1, 2 P30989, O95665 50 - 6 WD repeat-containing protein 5, 5B P61964, Q86VZ2 40 45 7 Coagulation factor VII, IXa heavy chain P08709, P00740 40 40 8 Factor X light chain P00742 40 40 9 Complex (Integrin beta-3) P08514/P05106 35 50 10 E3 ubiquitin-protein ligase XIAP P98170 – 45 11 Baculoviral IAP repeat-containing protein 2, 3, 8 Q13490, Q13489, Q96P09 – 45 Probability on target was computed based on a cross-validation. They may therefore not represent the actual probability of success for any new molecule. Thevelein et al. 1995). T6P plays a critical role as a sensing molecule that promotes sugar fermentation and glucose repression in yeast (Vicente et al. 2018). Thus, trehalose diet by cancer patient may be necessary to trigger energy stress which will in turn open up cancer cell to the pathway of selenoprotein BthD. Isomaltose will be needed to mitigate against the diabetes signaling molecules that are associated with cancer cell proliferation. Isomaltose inhibit adenylosuccinate lyase which is a more proximal enzyme in the adenylosuccinate biosynthesis pathway, lowers S-AMP levels and impairs glucose- stimulated insulin secretion (Fatoki et al. 2018b), and may help to reduce the risks associated with obesity and type 2 diabetes (van Can et al. 2012). Previous studies haves established that phosphoglycerate kinase 1 (PGK1) and pyruvate kinase M2 (PKM2) the only two ATP-generating glycolytic enzymes, which function as protein kinases and play active roles in tumor development (Li et al. 2016a). Another study has shown that hepatocellular carcinoma (HCC) cells reduce the fructose metabolism rate, and involved a switch in expression from fructokinase C (KHK-C) to fructokinase A (KHK-A) (Li et al. 2016b), and in the process KHK-A enhanced nucleic acid synthesis for tumorigenesis (Li et al. 2016c), and also enhanced p62’s aggregation with Kelch- like ECH-associated protein 1 (Keap1) and nuclear factor erythroid 2-related factor 2 (Nrf2) activation (Xu et al. 2019). Nrf2 is located in the cytoplasm and guided by Keap1, but under oxidative stress Nrf2 moves to the nucleus, where it binds the antioxidant response element (ARE) and drives the expression of several downstream genes such as γ-glutamyl cysteine synthetase modifier subunit (GCLm), glutamate cysteine ligase catalytic subunit (GCLC), heme oxygenase-1 (HO-1) and NAD(P)H quinine oxidoreductase-1 (NQO1) (Shibata et al. 2008; Singh et al. 2008; Ding et al. 2010; Kansanen et al. 2013). A novel peptide activator of a key antioxidant gene transcription pathwayin the hippocampus, which disrupted the Nrf2–Keap1 interaction in global cerebral ischemia model has been reported (Tu et al. 2015). The Hippo signaling pathway plays a crucial role in cell proliferation, apoptosis, differentiation, and development. Transcriptional co-activators Yes-associated protein 1 (YAP) and WW domain- containing transcription regulator protein 1 (TAZ), are major effectors of the Hippo signaling pathway (Bae et al. 2017). They function as transcription factors along with TEAD (TEA domain family member) in the nucleus, which increases expression of such target genes as Ctgf, Cyr61, AXL, and Survivin (Bae et al. 2017). Study has shown that phosphofructokinase 1 (PFK1) mediates glucose-induced YAP- and TAZ-TEAD interactions (Enzo et al. 2015). Energy stress, as induced by culturing cells in glucose-free conditions, results in inhibition of YAP activity in mouse hepatocytes in vivo as demonstrated by starvation/re-feeding experiments (Wang et al. 2015). In recent time, small peptides having anticancer properties have emerged as a potential Nova Biotechnol Chim (2020) 19(1): 37-51 48 Table 8. Pharmacokinetics of CRSUR Peptide from Selenoprotein BthD. S/N Parameters CRSUR Peptide Linear Head-to-tail bond 1 Molecular Weight [g.mol-1] 669.64 651.62 2 Heavy Atoms (HA) 41 40 3 Molar Refractivity 151.53 167.52 4 Total Polar Surface Area (Å2) 362.55 328.33 5 Lipophilicity Consensus LogP -3.96 -3.73 6 Water Solubility ESOL Class Highly soluble Very soluble 7 Gastrointestinal Absorption Low Low 8 Blood Brain Barrier (BBB) Permeant No No 9 P-glycoprotein Substrate No No 10 Cytochrome P450s Inhibitor No No 11 Skin permeation log Kp [cm.s-1] -15.29 -12.84 12 Lipinski Violation 3 3 13 Bioavailability Score 0.17 0.17 14 Synthetic Accessibility 5.62 6.40 alternative approach for cancer therapy (Thundimadathil et al. 2012). Anticancer peptides (ACPs) are small (5 – 30 amino acids) peptides, often derived from antimicrobial peptides (AMPs) and are cationic in nature (Tyagi et al. 2013), while cell-penetrating peptides (CPPs) are small peptides that have unique inherent ability to directly enter cells without significantly damaging the cell membrane (Wei et al. 2017). In this study, the selenocysteine-containing peptide CRSUR was identified and further investigated among all peptides obtained. The results show that cyclic peptide CRSUR will be a good anticancer agent through E3 ubiquitin-protein ligase regulating NF- kappa-B signalling by unleashes cell for spontaneous formation of the ripoptosome, a large multi-protein complex that has the capability to kill cancer cells in a caspase-dependent and caspase- independent manner (Bertrand et al. 2011). Conclusions Low calories diets can cause significant reduction in tumor incidence and tumor growth, and the mechanistic links between diet and cancer which has remain poorly understood (Warr et al. 2018), has been unravelled for application in human health through this in-silico study. We have shown that selenoprotein BthD can be good antioxidant supplement alone or together with the whole fruit juice of V. vinifera, not only against cancer but also virus infection and for overall human health (Moghadaszadeh and Beggs 2006). Thus, it is possible to build on the understanding of the antioxidant/anticancer potential of BthD by investigating new synthetic peptides from the conserved regions. Further study will be to evaluate anticancer potential of optimized peptides of selenoprotein BthD consisting of 5 – 10 amino acid residues; and also investigate the Sec- containing disaccharides as novel anticancer compounds. 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