Broad-Spectrum Antiviral Activity of an Ankyrin Repeat Protein on Viral Assembly against Chimeric NL4-3 Viruses Carrying Gag/PR Derived from Circulating Strains among Northern Thai Patients

Sakkhachornphop, Supachai; Hadpech, Sudarat; Wisitponchai, Tanchanok; Panto, Chansunee; Kantamala, Doungnapa; Utaipat, Utaiwan; Praparattanapan, Jutarat; Kotarathitithum, Wilai; Taejaroenkul, Sineenart; Yasamut, Umpa; Chupradit, Koollawat; Moonmuang, Sutpirat; Lee, Vannajan Sanghiran; Suparatpinyo, Khuanchai; Tayapiwatana, Chatchai

doi:10.3390/v10110625

Cited by 6 publications

(8 citation statements)

References 45 publications

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“…Previously, our group also reported that protein scaffolds including ankyrin repeat protein, zinc finger protein, and αRep protein could be used as intracellular antiviral molecules, and also demonstrated their therapeutic potential as HIV-1 inhibitors [17,[28][29][30][31].…”

Section: Discussionmentioning

confidence: 97%

Occupation of a thermoresistant-scaffold (αRep) at SP1-NC cleavage site disturbs the function of HIV-1 protease

et al. 2020

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HIV-1 nucleocapsid (NC) becomes an attractive target for the development of novel anti-HIV-1 agents. Discovering of non-antibody scaffolds that disrupt the function of NC will be a potential aspect for disturbing viral maturation process. Correspondingly, we explored the specific binding site of the thermoresistant-scaffold protein, αRep9A8 which formerly demonstrated the inhibitory effect on HIV-1 replication. The portion of Gag, CA21-SP1-NC has been used as a template for designing nine overlapping peptides (P4–P12). The P9 peptide showed the strongest binding activity followed by P8 and P12 respectively. The amino acid sequences on those peptides resemble the N-terminal domain of the NC proximity to the SP1-NC initial cleavage site and across the conserved CCHC zinc finger 1 (ZF1) of NC. The interaction KD between αRep9A8 with its target was 224.9 ± 57.4 nM. Consequently, αRep9A8 demonstrated the interference of the HIV-1 protease function by hindering a protease cleavage site. The released NC product from CA21-SP1-NC was diminished. The present study provided an additional information of αRep9A8 function in interfering of viral maturation processes resulting in the decremental efficiency of viral infectivity.

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Section: Discussionmentioning

confidence: 97%

Occupation of a thermoresistant-scaffold (αRep) at SP1-NC cleavage site disturbs the function of HIV-1 protease

et al. 2020

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“…SupT1 cells that stably express an artificial ankyrin repeat molecule (AnkGAG1D4) binding to the human immunodeficiency virus (HIV) Gag polyprotein showed a reduced permissiveness to HIV-1 infection [ 34 ]. Subsequent studies investigated the molecular mechanisms whereby AnkGAG1D4 inhibited HIV replication, showing that the intracellular expression of AnkGAG1D4 affected the late stages of virus production [ 66 , 67 , 68 , 69 ]. Despite this as a promising strategy, intracellular DARPin delivery may limit in vivo administration of the molecule.…”

Section: Viral Applicationsmentioning

confidence: 99%

Designed Ankyrin Repeat Proteins: A New Class of Viral Entry Inhibitors

Walser

Mayor

Rothenberger

2022

Viruses

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Designed ankyrin repeat proteins (DARPins) are engineered proteins comprising consensus designed ankyrin repeats as scaffold. Tightly packed repeats form a continuous hydrophobic core and a large groove-like solvent-accessible surface that creates a binding surface. DARPin domains recognizing a target of interest with high specificity and affinity can be generated using a synthetic combinatorial library and in vitro selection methods. They can be linked together in a single molecule to build multispecific and multifunctional proteins without affecting expression or function. The modular architecture of DARPins offers unprecedented possibilities of design and opens avenues for innovative antiviral strategies.

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“…Certain proteins are currently attracting attention for the interference of HIV-1 replication. The potential of designed ankyrin-repeat proteins (DARPins) has been proposed as an alternative treatment to monoclonal antibodies [ 1 , 2 , 3 ]. Ank GAG 1D4 is an intracellular anti-HIV-1 DARPin that specifically targets HIV-1 capsid CAp24 [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

Dimeric Ankyrin with Inverted Module Promotes Bifunctional Property in Capturing Capsid to Impede HIV-1 Replication

Juntit

Sornsuwan

Wisitponchai

et al. 2023

IJMS

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Several anti-HIV scaffolds have been proposed as complementary treatments to highly active antiretroviral therapy. AnkGAG1D4, a designed ankyrin repeat protein, formerly demonstrated anti-HIV-1 replication by interfering with HIV-1 Gag polymerization. However, the improvement of the effectiveness was considered. Recently, the dimeric molecules of AnkGAG1D4 were accomplished in enhancing the binding activity against HIV-1 capsid (CAp24). In this study, the interaction of CAp24 against the dimer conformations was elucidated to elaborate the bifunctional property. The accessibility of the ankyrin binding domains was inspected by bio-layer interferometry. By inverting the second module of dimeric ankyrin (AnkGAG1D4NC-CN), the CAp24 interaction KD was significantly reduced. This reflects the capability of AnkGAG1D4NC-CN in simultaneously capturing CAp24. On the contrary, the binding activity of dimeric AnkGAG1D4NC-NC was indistinguishable from the monomeric AnkGAG1D4. The bifunctional property of AnkGAG1D4NC-CN was subsequently confirmed in the secondary reaction with additional p17p24. This data correlates with the MD simulation, which suggested the flexibility of the AnkGAG1D4NC-CN structure. The CAp24 capturing capacity was influenced by the distance of the AnkGAG1D4 binding domains to introduce the avidity mode of AnkGAG1D4NC-CN. Consequently, AnkGAG1D4NC-CN showed superior potency in interfering with HIV-1 NL4-3 WT and HIV-1 NL4-3 MIRCAI201V replication than AnkGAG1D4NC-NC and an affinity improved AnkGAG1D4-S45Y.

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Broad-Spectrum Antiviral Activity of an Ankyrin Repeat Protein on Viral Assembly against Chimeric NL4-3 Viruses Carrying Gag/PR Derived from Circulating Strains among Northern Thai Patients

Cited by 6 publications

References 45 publications

Occupation of a thermoresistant-scaffold (αRep) at SP1-NC cleavage site disturbs the function of HIV-1 protease

Occupation of a thermoresistant-scaffold (αRep) at SP1-NC cleavage site disturbs the function of HIV-1 protease

Designed Ankyrin Repeat Proteins: A New Class of Viral Entry Inhibitors

Dimeric Ankyrin with Inverted Module Promotes Bifunctional Property in Capturing Capsid to Impede HIV-1 Replication

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