A phage display‐selected peptide inhibitor of <scp><i>A</i></scp><i>grobacterium vitis</i> polygalacturonase

Warren, Jeremy G.; Kasun, George W.; Leonard, Takara; Kirkpatrick, Bruce C.

doi:10.1111/mpp.12293

Cited by 4 publications

(3 citation statements)

References 34 publications

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“…Moreover, they have been explored as therapeutic agents, such as vaccines, [20] immunotherapy agents, [21] and inhibitors of a target protein. [22][23][24][25] For example, the world's best-selling drug, HUMIRA, was discovered using the phage display technology. We recently developed a novel biopanning strategy to discover small peptides that can specifically bind to programmed death-ligand 1 (PD-L1) and block its interaction with the programmed cell death 1 (PD-1).…”

Section: Introductionmentioning

confidence: 99%

Discovery of Small Anti‐ACE2 Peptides to Inhibit SARS‐CoV‐2 Infectivity

Adhikary

Kandel

Mamani

et al. 2021

Advanced Therapeutics

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COVID‐19 is caused by the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), which infects host cells by binding its viral spike protein receptor‐binding domain (RBD) to the angiotensin converting enzyme 2 (ACE2) on host cells. Blocking the SARS‐CoV‐2‐RBD/ACE2 interaction is, therefore, a potential strategy to inhibit viral infections. Using a novel biopanning strategy, a small anti‐ACE2 peptide is discovered, which shows high affinity and specificity to human ACE2. It blocks not only the SARS‐CoV‐2‐RBD/ACE2 interaction but also the SARS‐CoV‐1‐RBD/ACE2 interaction. Moreover, it inhibits SARS‐CoV‐2 infection in Vero‐E6 cells. The peptide shows negligible cytotoxicity in Vero‐E6 cells and Huh7 cells. In vivo short‐term lung toxicity study also demonstrates a good safety of the peptide after intratracheal administration. The anti‐ACE2 peptide can be potentially used as a prophylactic or therapeutic agent for SARS‐CoV‐2 or other ACE2‐mediated viruses. The strategy used in this study also provides a fast‐track platform to discover other antiviral peptides, which will prepare the world for future pandemics.

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Section: Introductionmentioning

confidence: 99%

Discovery of Small Anti‐ACE2 Peptides to Inhibit SARS‐CoV‐2 Infectivity

Adhikary

Kandel

Mamani

et al. 2021

Advanced Therapeutics

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“…vitis polygalacturonase can be readily produced in recombinant E . coli expression systems [ 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

Insights into the Activity and Substrate Binding of Xylella fastidiosa Polygalacturonase by Modification of a Unique QMK Amino Acid Motif Using Protein Chimeras

2015

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Polygalacturonases (EC 3.2.1.15) catalyze the random hydrolysis of 1, 4-alpha-D-galactosiduronic linkages in pectate and other galacturonans. Xylella fastidiosa possesses a single polygalacturonase gene, pglA (PD1485), and X. fastidiosa mutants deficient in the production of polygalacturonase are non-pathogenic and show a compromised ability to systemically infect grapevines. These results suggested that grapevines expressing sufficient amounts of an inhibitor of X. fastidiosa polygalacturonase might be protected from disease. Previous work in our laboratory and others have tried without success to produce soluble active X. fastidiosa polygalacturonase for use in inhibition assays. In this study, we created two enzymatically active X. fastidiosa / A. vitis polygalacturonase chimeras, AX1A and AX2A to explore the functionality of X. fastidiosa polygalacturonase in vitro. The AX1A chimera was constructed to specifically test if recombinant chimeric protein, produced in Escherichia coli, is soluble and if the X. fastidiosa polygalacturonase catalytic amino acids are able to hydrolyze polygalacturonic acid. The AX2A chimera was constructed to evaluate the ability of a unique QMK motif of X. fastidiosa polygalacturonase, most polygalacturonases have a R(I/L)K motif, to bind to and allow the hydrolysis of polygalacturonic acid. Furthermore, the AX2A chimera was also used to explore what effect modification of the QMK motif of X. fastidiosa polygalacturonase to a conserved RIK motif has on enzymatic activity. These experiments showed that both the AX1A and AX2A polygalacturonase chimeras were soluble and able to hydrolyze the polygalacturonic acid substrate. Additionally, the modification of the QMK motif to the conserved RIK motif eliminated hydrolytic activity, suggesting that the QMK motif is important for the activity of X. fastidiosa polygalacturonase. This result suggests X. fastidiosa polygalacturonase may preferentially hydrolyze a different pectic substrate or, alternatively, it has a different mechanism of substrate binding than other polygalacturonases characterized to date.

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“…Of these enzymes, polygalacturonase (PG) is one of the most important virulence factors. Plants also produce PGs during the ripening process to soften and sweeten fruits10. According to the sequence homology and three-dimensional structures, PGs are confined to family 28 of the glycoside hydrolases (GH; http://www.cazy.org/GH28.html)11.…”

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confidence: 99%

Probing the role of cation-π interaction in the thermotolerance and catalytic performance of endo-polygalacturonases

et al. 2016

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Understanding the dynamics of the key pectinase, polygalacturonase, and improving its thermotolerance and catalytic efficiency are of importance for the cost-competitive bioconversion of pectic materials. By combining structure analysis and molecular dynamics (MD) simulations, eight mutagenesis sites having the potential to form cation-π interactions were identified in the widely used fungal endo-polygalacturonase PG63. In comparison to the wild-type, three single mutants H58Y, T71Y and T304Y showed improved thermostability (the apparent Tms increased by 0.6−3.9 °C) and catalytic efficiency (by up to 32-fold). Chromatogram analysis of the hydrolysis products indicated that a larger amount of shorter sugars were released from the polygalacturonic acid by these three mutants than by the wild-type. MD analysis of the enzyme-substrate complexes illustrated that the mutants with introduced cation-π interaction have modified conformations of catalytic crevice, which provide an enviable environment for the catalytic process. Moreover, the lower plasticity of T3 loop 2 at the edge of the subsite tunnel appears to recruit the reducing ends of oligogalacturonide into the active site tunnel and initiates new hydrolysis reactions. This study demonstrates the importance of cation-π interaction in protein conformation and provides a realistic strategy to enhance the thermotolerance and catalytic performance of endo-polygalacturonases.

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A phage display‐selected peptide inhibitor of Agrobacterium vitis polygalacturonase

Cited by 4 publications

References 34 publications

Discovery of Small Anti‐ACE2 Peptides to Inhibit SARS‐CoV‐2 Infectivity

Discovery of Small Anti‐ACE2 Peptides to Inhibit SARS‐CoV‐2 Infectivity

Insights into the Activity and Substrate Binding of Xylella fastidiosa Polygalacturonase by Modification of a Unique QMK Amino Acid Motif Using Protein Chimeras

Probing the role of cation-π interaction in the thermotolerance and catalytic performance of endo-polygalacturonases

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