Identification and Quantification of DPP-IV-Inhibitory Peptides from Hydrolyzed-Rapeseed-Protein-Derived Napin with Analysis of the Interactions between Key Residues and Protein Domains

Xu, Feiran; Yao, Yijun; Xu, Xiaoying; Wang, Mei; Pan, Mengmeng; Ji, Shengyang; Wu, Jin; Jiang, Donglei; Ju, Xingrong; Wang, Lifeng

doi:10.1021/acs.jafc.9b01069

Cited by 68 publications

(72 citation statements)

References 49 publications

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“…Napin migrated as a doublet in the range of 5-10 kDa, whereas cruciferin migrated as double triplets, the upper one with protein bands in the range of 25-35 kDa and the lower one with bands in the range of 15-20 kDa (Figure 1). The protein electrophoretic profile matches those from earlier reports (Höglund et al, 1992;Wanasundara, 2011;Stone et al, 2014;Akbari and Wu, 2015;Perera et al, 2016;Joehnke et al, 2019;Xu et al, 2019;Rahman et al, 2020) ( Supplementary Figure 2A).…”

Section: Protein Characterizationsupporting

confidence: 86%

In Silico, Molecular Docking and In Vitro Antimicrobial Activity of the Major Rapeseed Seed Storage Proteins

et al. 2020

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Background: In addition to their use as an edible oil and condiment crop, mustard and rapeseed (Brassica napus L., B. juncea (L.) Czern., B. nigra (L.) W.D.J.Koch, B. rapa L. and Sinapis alba L.) have been commonly used in traditional medicine for relieving pain, coughs and treating infections. The seeds contain high amounts of oil, while the remaining byproduct meal after oil extraction, about 40% of seed dry weight, has a low value despite its high protein-content (~85%). The seed storage proteins (SSP) 2S albumin-type napin and 12S globulin-type cruciferin are the two predominant proteins in the seeds and show potential for value adding to the waste stream; however, information on their biological activities is scarce. In this study, purified napin and cruciferin were tested using in silico, molecular docking, and in vitro approaches for their bioactivity as antimicrobial peptides. Materials and Methods: The 3D-structure of 2S albumin and 12S globulin storage proteins from B. napus were investigated to predict antimicrobial activity employing an antimicrobial peptide database survey. To gain deeper insights into the potential antimicrobial activity of these SSP, in silico molecular docking was performed. The purified B. napus cruciferin and napin were then tested against both Gram-positive and Gram-negative bacteria for in vitro antimicrobial activity by disc diffusion and microdilution antimicrobial susceptibility testing.

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Section: Protein Characterizationsupporting

confidence: 86%

In Silico, Molecular Docking and In Vitro Antimicrobial Activity of the Major Rapeseed Seed Storage Proteins

et al. 2020

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“…In our previous study (Xu et al, 2019), the octapeptide ELHQEEPL (IC 50 = 78.46 ± 4.94 μM) was confirmed to demonstrate an excellent DPP-IV inhibitory DPP-IV using in vitro enzymatic assay.…”

Section: Introductionmentioning

confidence: 75%

“…The main objective of the study was to assess whether ELHQEEPL could maintain the same DPP-IV inhibitory activity in Caco-2 cell monolayers model as demonstrated in our previous results (Xu et al, 2019). Well-differentiated Caco-2 cells possess a microvilli structure on the apical surface, with potential polarity related to the basolateral side (Marques et al, 2011;Pshezhetsky et al, 2007), and have been reported to contain many brush border proteases (BBPs),…”

Section: Discussionmentioning

confidence: 94%

“…Gly-Pro-p-nitroanilide as described by our previous publication (Xu et al, 2019) where A 405 (sample) is the absorbance at 405 nm (OD 405 ) in the presence of DPP-IV, the reaction substrate (Gly-Pro-p-nitroanilide), and a peptide sample; A 405 (sample/control) is the OD 405 when DPP-IV is substituted with Tris-HCl buffer (100 mM, pH 8.0) in the sample assay; A 405 (negative/reaction) is the OD 405 of DPP-IV and reaction substrate with no inhibitor; and A 405 (negative/control) is the OD 405 of the assay with no DPP-IV or inhibitor.…”

Section: In Vitro Dpp-iv Activity Inhibition Assay In Chemical Enzymentioning

confidence: 99%

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Assessment of the DPP‐IV inhibitory activity of a novel octapeptide derived from rapeseed using Caco‐2 cell monolayers and molecular docking analysis

Mejı́a

Chen

et al. 2020

J. Food Biochem.

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The Octapeptide ELHQEEPL, which was identified from the rapeseed protein napin showed prominent Dipeptidyl peptidase-IV (DPP-IV) inhibitory activity. The objective of this study was to investigate the DPP-IV inhibitory activity and transepithelial transport of ELHQEEPL in an approaching intestinal condition using Caco-2 cell monolayers. ELHQEEPL and its degraded fragments EL, HQEEP, and methylated ELHQEEPL were transported across Caco-2 cell monolayers through different pathways. Compared with the nonbiological enzyme inhibition test, the in vitro experiment on Caco-2 cell monolayers showed that the IC 50 value of DPP-IV inhibition increased by 43.11% for ELHQEEPL. There was no significant change in DPP-IV gene expression in the Caco-2 cell monolayers upon treatment with ELHQEEPL. Furthermore, molecular docking predicted that the weaker binding between inhibitory peptide and enzyme for the degradation products from ELHQEEPL during transepithelial transport greatly limited its role in inhibiting DPP-IV. Practical applications The DPP-IV inhibitory activity of ELHQEEPL was confirmed using Caco-2 cell monolayers as a novel assessment tool, although its potency was reduced by metabolic degradation. In general, this study reported the use of Caco-2 cell monolayers as a tool for comprehensively studying peptides as sources of DPP-IV inhibitors. A Caco-2 cell-based approach with molecular docking can be adapted for the investigation of intestinal absorption and activity attenuation of food peptides being considered for enzymatic action. Moreover, since the Caco-2 cells express a wide range of enzymes, this method can be used for screening for other active food peptides such as for the

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“…Adding two more amino acids at the C terminus changes the DPP-IV inhibitory action and the modification of targeting regions and binding sites on DPP-IV. Until now, most food protein-derived peptides were reported to be noncompetitive inhibitors or uncompetitive inhibitors [ 19 , 34 , 39 , 40 , 41 ]. Therefore, this research recommends an innovative concept for developing DPP-IV inhibitory peptides for functional food industry.…”

Section: Resultsmentioning

confidence: 99%

Characterization of Novel Dipeptidyl Peptidase-IV Inhibitory Peptides from Soft-Shelled Turtle Yolk Hydrolysate Using Orthogonal Bioassay-Guided Fractionations Coupled with In Vitro and In Silico Study

et al. 2020

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Five novel peptides (LPLF, WLQL, LPSW, VPGLAL, and LVGLPL) bearing dipeptidyl peptidase IV (DPP-IV) inhibitory activities were identified from the gastrointestinal enzymatic hydrolysate of soft-shelled turtle yolk (SSTY) proteins. Peptides were isolated separately using reversed-phase (RP) chromatography in parallel with off-line strong cation exchange (SCX) chromatography followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis to determine sequences. Among these peptides, LPSW showed the highest DPP-IV inhibitory activity with an IC50 value of 269.7 ± 15.91 µM. The results of the pre-incubation experiment and the kinetic study of these peptides indicated that WLQL is a true inhibitor and its inhibition toward DPP-IV is of an uncompetitive model, while LPLF, LPSW, and VPGLAL are real-substrates and competitive inhibitors against DPP-IV. The DPP-IV inhibitory peptides derived from SSTY hydrolysate in study are promising in the management of hyperglycemia in Type 2 diabetes.

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Identification and Quantification of DPP-IV-Inhibitory Peptides from Hydrolyzed-Rapeseed-Protein-Derived Napin with Analysis of the Interactions between Key Residues and Protein Domains

Cited by 68 publications

References 49 publications

In Silico, Molecular Docking and In Vitro Antimicrobial Activity of the Major Rapeseed Seed Storage Proteins

In Silico, Molecular Docking and In Vitro Antimicrobial Activity of the Major Rapeseed Seed Storage Proteins

Assessment of the DPP‐IV inhibitory activity of a novel octapeptide derived from rapeseed using Caco‐2 cell monolayers and molecular docking analysis

Characterization of Novel Dipeptidyl Peptidase-IV Inhibitory Peptides from Soft-Shelled Turtle Yolk Hydrolysate Using Orthogonal Bioassay-Guided Fractionations Coupled with In Vitro and In Silico Study

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