Identification and characterization of a wolfberry carboxypeptidase inhibitor from Lycium barbarum

Huang, Jiayi; Wong, Ka Ho; Tan, Wei; Tay, Stephanie; Wang, Shujing; Tam, James P.

doi:10.1016/j.foodchem.2021.129338

Cited by 12 publications

(8 citation statements)

References 39 publications

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“…Another study, which characterized a new CPI named β-lybatide from Lycium barbarum , also constructed a phylogenetic tree of protease inhibitors. The resulting clade corresponding to plant CPIs exhibited similar relationships as described in this study, wherein sequences from the same genus showed greater conservation and closer phylogenetic ties ( Huang et al, 2021 ). This finding further supports the evolutionary relationships described in this study, providing additional evidence for the relatedness and diversification of CPIs within the Solanaceae family.…”

Section: Discussionsupporting

confidence: 63%

Carboxypeptidase inhibitors from Solanaceae as a new subclass of pathogenesis related peptide aiming biotechnological targets for plant defense

Gomes,

Espósito,

Baracat-Pereira

2023

Front. Mol. Biosci.

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Background: Plant protease inhibitors play a crucial role in inhibiting proteases produced by phytopathogens and exhibiting inhibitory effects on nematodes, fungi, and insects, making them promising candidates for crop protection. Specifically, carboxypeptidase inhibitors, a subset of proteinase inhibitors, have been extensively studied in potato and tomato of Solanaceae plant family. However, further research is needed to fully understand the functions and biotechnological potential of those inhibitors in plants. This work aimed to in silico characterize carboxypeptidase inhibitors from Solanaceae as potential antimicrobial and defense agents focused on biotechnological targets.Methods: The methodology employed involved search in UniProt, PDB, KNOTTIN, NCBI, and MEROPS databases for solanaceous carboxypeptidase inhibitors, phylogenetic relationships and conservation patterns analyzes using MEGA-X software and Clustal Omega/MView tools, physicochemical properties and antimicrobial potential prediction using ProtParam, ToxinPred, iAMPred, and APD3 tools, and structural features prediction using PSIPRED.Results and discussion: A systematic literature search was conducted to identify relevant studies on Solanaceae carboxypeptidase inhibitors and their activities against pathogens. The selected studies were reviewed and the main findings compiled. The characterization of Solanaceae carboxypeptidase inhibitors proposed for the first time the global sequence consensus motif CXXXCXXXXDCXXXXXCXXC, shedding light on carboxypeptidase inhibitors distribution, sequence variability, and conservation patterns. Phylogenetic analysis showed evolutionary relationships within the Solanaceae family, particularly in Capsicum, Nicotiana, and Solanum genera. Physicochemical characteristics of those peptides indicated their similarity to antimicrobial peptides. Predicted secondary structures exhibited variations, suggesting a broad spectrum of action, and studies had been demonstrated their activities against various pathogens.Conclusion: Carboxypeptidase inhibitors are being proposed here as a new subclass of PR-6 pathogenesis-related proteins, which will aid in a focused understanding of their functional roles in plant defense mechanisms. These findings confirm the Solanaceae carboxypeptidase inhibitors potential as defense agents and highlight opportunities for their biotechnological applications in pathogen control.

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Section: Discussionsupporting

confidence: 63%

Carboxypeptidase inhibitors from Solanaceae as a new subclass of pathogenesis related peptide aiming biotechnological targets for plant defense

Gomes,

Espósito,

Baracat-Pereira

2023

Front. Mol. Biosci.

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“…Apart from PCI, other well‐studied inhibitors include Lycium barbarum carboxypeptidase inhibitor (WCI), 30 the tick carboxypeptidase inhibitor (TCI), 22 and the leech carboxypeptidase inhibitor (LCI) 31 . WCI is the closest to PCI in terms of size (4 kDa) and amino acid residue conservation (64% identity; Figure S3).…”

Section: Resultsmentioning

confidence: 99%

Structure of Aedes aegypti carboxypeptidase B1‐inhibitor complex uncover the disparity between mosquito and non‐mosquito insect carboxypeptidase inhibition mechanism

et al. 2021

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Metallocarboxypeptidases (MCPs) in the mosquito midgut play crucial roles in infection, as well as in mosquito dietary digestion, reproduction, and development. MCPs are also part of the digestive system of plant‐feeding insects, representing key targets for inhibitor development against mosquitoes/mosquito‐borne pathogens or as antifeedant molecules against plant‐feeding insects. Notably, some non‐mosquito insect B‐type MCPs are primarily insensitive to plant protease inhibitors (PPIs) such as the potato carboxypeptidase inhibitor (PCI; MW 4 kDa), an inhibitor explored for cancer treatment and insecticide design. Here, we report the crystal structure of Aedes aegypti carboxypeptidase‐B1 (CPBAe1)‐PCI complex and compared the binding with that of PCI‐insensitive CPBs. We show that PCI accommodation is determined by key differences in the active‐site regions of MCPs. In particular, the loop regions α6‐α7 (Leu242‐Ser250) and β8‐α8 (Pro269‐Pro280) of CPBAe1 are replaced by α‐helices in PCI‐insensitive insect Helicoverpa zea CPBHz. These α‐helices protrude into the active‐site pocket of CPBHz, restricting PCI insertion and rendering the enzyme insensitive. We further compared our structure with the only other PCI complex available, bovine CPA1‐PCI. The potency of PCI against CPBAe1 (Ki = 14.7 nM) is marginally less than that of bovine CPA1 (Ki = 5 nM). Structurally, the above loop regions that accommodate PCI binding in CPBAe1 are similar to that of bovine CPA1, although observed changes in proteases residues that interact with PCI could account for the differences in affinity. Our findings suggest that PCI sensitivity is largely dictated by structural interference, which broadens our understanding of carboxypeptidase inhibition as a mosquito population/parasite control strategy.

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“…43 C-(C�O) bond cleaved by carboxypeptidase (Figure 5): Carboxypeptidase is specifically able to remove C-terminal residues from peptides but can also hydrolyze longer peptide chains. 44 The formation of KK, C 16 H 35 N 5 O 2 Se "m/z 408.18722" was a result of the C-terminal methionine of the KKSe(M)R being cracked by the action of carboxypeptidase on the C-(C�O) bond. Rapid cleavage of the amide bond in KKSe(M)R (Figure 5): Amide bonds serve nontrivial biological functions, and their stability is facilitated by their intrinsic ability to form resonance structures.…”

Section: Simulated Processing Conditions Mediated the Pyrolysis-therm...mentioning

confidence: 99%

Se-Pair Search for Deciphering Selenium-Encoded Peptide and a Pyrolysis-Thermolysis Dietary Model for Minimizing Loss of “KKSe(M)R” during Processing

Wang,

Jia

2024

J. Agric. Food Chem.

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Dietary deficiency of selenium is a global health hazard. Supplementation of organic selenopeptides via food crops is a relatively safe approach. Selenopeptides with heterogeneous selenium-encoded isotopes or a poorly fragmented peptide backbone remain unidentified in site-specific selenoproteomic analysis. Herein, we developed the Se-Pair Search, a UniProtKB-FASTA-independent peptide-matching strategy, exploiting the fragmentation patterns of shared peptide backbones in selenopeptides to optimize spectral interpretation, along with developing new selenosite assignment schemes (steps 1–3) to standardize selenium-localization data reporting for the selenoproteome community and thereby facilitating the discovery of unexpected selenopeptides. Using selenium-biofortified rice under cooking, fermentation, and high-temperature and high-pressure processing conditions as a pyrolysis-thermolysis dietary model, we probed the single-molecule-level kinetic evolution of the novel selenopeptide “KKSe(M)R” with qual-quantitative information on graph-theory-oriented localization calculations, abundance patterns, activation energy, and rate constants at a selenoproteome-wide scale. We ground-truth-annotated thirteen pyrolysis-thermolysis products and inferred four pyrolysis-thermolysis pathways to characterize the formation reactivity of the main intermediate variables of KKSe(M)R and constructed an advanced probe-type ultrasound technique prior to pyrolysis-thermolysis conditions for minimizing loss of KKSe(M)R during processing. Importantly, we reveal the unappreciated pyro-excitation diversion of KKSe(M)R at pyrolysis-thermolysis time and temperature matrices. These findings provide pioneering theoretical guidance for controlling dietary selenium supplementation within the safety thresholds.

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Identification and characterization of a wolfberry carboxypeptidase inhibitor from Lycium barbarum

Cited by 12 publications

References 39 publications

Carboxypeptidase inhibitors from Solanaceae as a new subclass of pathogenesis related peptide aiming biotechnological targets for plant defense

Carboxypeptidase inhibitors from Solanaceae as a new subclass of pathogenesis related peptide aiming biotechnological targets for plant defense

Structure of Aedes aegypti carboxypeptidase B1‐inhibitor complex uncover the disparity between mosquito and non‐mosquito insect carboxypeptidase inhibition mechanism

Se-Pair Search for Deciphering Selenium-Encoded Peptide and a Pyrolysis-Thermolysis Dietary Model for Minimizing Loss of “KKSe(M)R” during Processing

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