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Identification of restriction endonuclease with potential ability to cleave the HSV-2 genome: inherent potential for biosynthetic versus live recombinant microbicides.

Wayengera M, Kajumbula H, Byarugaba W - Theor Biol Med Model (2008)

Bottom Line: Developing biomedical strategies for HSV-2 prevention is thus a central strategy in reducing global HIV-1 prevalence.In silico palindromics has a PPV of 99.5% for in situ REase activity (2) Two models detailing how the REase EcoRII may be applied in developing interventions against HSV-2 are presented: a nanoparticle for microbicide development and a "recombinant lactobacillus" expressing cell wall anchored receptor (truncated nectin-1) for HSV-2 plus EcoRII.Viral genome slicing by way of these bacterially- derived R-M enzymatic peptides may have therapeutic potential in HSV-2 infection; a cofactor for HIV-1 acquisition and transmission.

View Article: PubMed Central - HTML - PubMed

Affiliation: Restrizymes Biotherapeutics Uganda Limited, Kampala, Uganda. wmisaki@yahoo.com

ABSTRACT

Background: Herpes Simplex virus types 1 and 2 are enveloped viruses with a linear dsDNA genome of approximately 120-200 kb. Genital infection with HSV-2 has been denoted as a major risk factor for acquisition and transmission of HIV-1. Developing biomedical strategies for HSV-2 prevention is thus a central strategy in reducing global HIV-1 prevalence. This paper details the protocol for the isolation of restriction endunucleases (REases) with potent activity against the HSV-2 genome and models two biomedical interventions for preventing HSV-2.

Methods and results: Using the whole genome of HSV-2, 289 REases and the bioinformatics software Webcutter2; we searched for potential recognition sites by way of genome wide palindromics. REase application in HSV-2 biomedical therapy was modeled concomitantly. Of the 289 enzymes analyzed; 77(26.6%) had potential to cleave the HSV-2 genome in > 100 but < 400 sites; 69(23.9%) in > 400 but < 700 sites; and the 9(3.1%) enzymes: BmyI, Bsp1286I, Bst2UI, BstNI, BstOI, EcoRII, HgaI, MvaI, and SduI cleaved in more than 700 sites. But for the 4: PacI, PmeI, SmiI, SwaI that had no sign of activity on HSV-2 genomic DNA, all 130(45%) other enzymes cleaved < 100 times. In silico palindromics has a PPV of 99.5% for in situ REase activity (2) Two models detailing how the REase EcoRII may be applied in developing interventions against HSV-2 are presented: a nanoparticle for microbicide development and a "recombinant lactobacillus" expressing cell wall anchored receptor (truncated nectin-1) for HSV-2 plus EcoRII.

Conclusion: Viral genome slicing by way of these bacterially- derived R-M enzymatic peptides may have therapeutic potential in HSV-2 infection; a cofactor for HIV-1 acquisition and transmission.

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Related in: MedlinePlus

This figure attempts to model the molecular binding of EcoRII to nonoxynol-9 or C31G (savvy) through the polyester PLGA. A. N-9 and EcoRII PLGA loaded nanoparticles: Note the orientation of the hydrogen and hydroxyl ions in the glycolic and lactic acids monomers of PLGA towards the hydroxyl and hydrogen ions in the N-9 and the REase nanoparticles model. The underlined dots signify that it is unknown which, covalent or hydrogen, bonds are involved. B. C31G and EcoRII PLGA loaded nanoparticles. Note that the chemical structure of Savvy is has been abbreviated to C31G, but is [C14H29 N(CH3)2O]A- + [C16H33 N (CH3)2CH2COO]B-.
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Figure 4: This figure attempts to model the molecular binding of EcoRII to nonoxynol-9 or C31G (savvy) through the polyester PLGA. A. N-9 and EcoRII PLGA loaded nanoparticles: Note the orientation of the hydrogen and hydroxyl ions in the glycolic and lactic acids monomers of PLGA towards the hydroxyl and hydrogen ions in the N-9 and the REase nanoparticles model. The underlined dots signify that it is unknown which, covalent or hydrogen, bonds are involved. B. C31G and EcoRII PLGA loaded nanoparticles. Note that the chemical structure of Savvy is has been abbreviated to C31G, but is [C14H29 N(CH3)2O]A- + [C16H33 N (CH3)2CH2COO]B-.

Mentions: For the purposes of achieving conjugation and chemical binding between either Savvy or Nonoxynol-9) and EcoRII, we further hypothesized that the aliphatic polyester poly(lactic-co-glycolic acid) (PLGA) may suffice [35]. PLGA is a copolymer that is synthesized by random ring-opening co-polymerization of two different monomers, the cyclic dimers (1,4-dioxane-2,5-diones) of glycolic acid and lactic acid on either tin (II) 2-ethylhexanoate, tin(II) alkoxides, or aluminum isopropoxide as catalysts. Owing to its wide solubility, bio-degradability and compatibility, PLGA is used in drug delivery by the formation of nanoparticles [36]. A simplified chemical structure of PLGA is shown in Figure 3. We finally derived a likely chemical structure of a single molecule of the nanoparticles: 1) nano-N-9-EcoRII and Nano-C31G-EcoRII. Both Theses model nanoparticle structures are shown in Figure 4. We believe that such nanoparticles may be synthesized practically using a two-step emulsion of EcoRII in PLGA followed by addition of N-9 or C31G rather than polyacrylic acid (PAA) as described elsewhere [35]. Note that it has been assumed that only a single molecule of EcoRII, C31G or N-9 and PLGA will form the nanoparticle, although practically speaking, the relative proportions of the constituent molecules may vary.


Identification of restriction endonuclease with potential ability to cleave the HSV-2 genome: inherent potential for biosynthetic versus live recombinant microbicides.

Wayengera M, Kajumbula H, Byarugaba W - Theor Biol Med Model (2008)

This figure attempts to model the molecular binding of EcoRII to nonoxynol-9 or C31G (savvy) through the polyester PLGA. A. N-9 and EcoRII PLGA loaded nanoparticles: Note the orientation of the hydrogen and hydroxyl ions in the glycolic and lactic acids monomers of PLGA towards the hydroxyl and hydrogen ions in the N-9 and the REase nanoparticles model. The underlined dots signify that it is unknown which, covalent or hydrogen, bonds are involved. B. C31G and EcoRII PLGA loaded nanoparticles. Note that the chemical structure of Savvy is has been abbreviated to C31G, but is [C14H29 N(CH3)2O]A- + [C16H33 N (CH3)2CH2COO]B-.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2526989&req=5

Figure 4: This figure attempts to model the molecular binding of EcoRII to nonoxynol-9 or C31G (savvy) through the polyester PLGA. A. N-9 and EcoRII PLGA loaded nanoparticles: Note the orientation of the hydrogen and hydroxyl ions in the glycolic and lactic acids monomers of PLGA towards the hydroxyl and hydrogen ions in the N-9 and the REase nanoparticles model. The underlined dots signify that it is unknown which, covalent or hydrogen, bonds are involved. B. C31G and EcoRII PLGA loaded nanoparticles. Note that the chemical structure of Savvy is has been abbreviated to C31G, but is [C14H29 N(CH3)2O]A- + [C16H33 N (CH3)2CH2COO]B-.
Mentions: For the purposes of achieving conjugation and chemical binding between either Savvy or Nonoxynol-9) and EcoRII, we further hypothesized that the aliphatic polyester poly(lactic-co-glycolic acid) (PLGA) may suffice [35]. PLGA is a copolymer that is synthesized by random ring-opening co-polymerization of two different monomers, the cyclic dimers (1,4-dioxane-2,5-diones) of glycolic acid and lactic acid on either tin (II) 2-ethylhexanoate, tin(II) alkoxides, or aluminum isopropoxide as catalysts. Owing to its wide solubility, bio-degradability and compatibility, PLGA is used in drug delivery by the formation of nanoparticles [36]. A simplified chemical structure of PLGA is shown in Figure 3. We finally derived a likely chemical structure of a single molecule of the nanoparticles: 1) nano-N-9-EcoRII and Nano-C31G-EcoRII. Both Theses model nanoparticle structures are shown in Figure 4. We believe that such nanoparticles may be synthesized practically using a two-step emulsion of EcoRII in PLGA followed by addition of N-9 or C31G rather than polyacrylic acid (PAA) as described elsewhere [35]. Note that it has been assumed that only a single molecule of EcoRII, C31G or N-9 and PLGA will form the nanoparticle, although practically speaking, the relative proportions of the constituent molecules may vary.

Bottom Line: Developing biomedical strategies for HSV-2 prevention is thus a central strategy in reducing global HIV-1 prevalence.In silico palindromics has a PPV of 99.5% for in situ REase activity (2) Two models detailing how the REase EcoRII may be applied in developing interventions against HSV-2 are presented: a nanoparticle for microbicide development and a "recombinant lactobacillus" expressing cell wall anchored receptor (truncated nectin-1) for HSV-2 plus EcoRII.Viral genome slicing by way of these bacterially- derived R-M enzymatic peptides may have therapeutic potential in HSV-2 infection; a cofactor for HIV-1 acquisition and transmission.

View Article: PubMed Central - HTML - PubMed

Affiliation: Restrizymes Biotherapeutics Uganda Limited, Kampala, Uganda. wmisaki@yahoo.com

ABSTRACT

Background: Herpes Simplex virus types 1 and 2 are enveloped viruses with a linear dsDNA genome of approximately 120-200 kb. Genital infection with HSV-2 has been denoted as a major risk factor for acquisition and transmission of HIV-1. Developing biomedical strategies for HSV-2 prevention is thus a central strategy in reducing global HIV-1 prevalence. This paper details the protocol for the isolation of restriction endunucleases (REases) with potent activity against the HSV-2 genome and models two biomedical interventions for preventing HSV-2.

Methods and results: Using the whole genome of HSV-2, 289 REases and the bioinformatics software Webcutter2; we searched for potential recognition sites by way of genome wide palindromics. REase application in HSV-2 biomedical therapy was modeled concomitantly. Of the 289 enzymes analyzed; 77(26.6%) had potential to cleave the HSV-2 genome in > 100 but < 400 sites; 69(23.9%) in > 400 but < 700 sites; and the 9(3.1%) enzymes: BmyI, Bsp1286I, Bst2UI, BstNI, BstOI, EcoRII, HgaI, MvaI, and SduI cleaved in more than 700 sites. But for the 4: PacI, PmeI, SmiI, SwaI that had no sign of activity on HSV-2 genomic DNA, all 130(45%) other enzymes cleaved < 100 times. In silico palindromics has a PPV of 99.5% for in situ REase activity (2) Two models detailing how the REase EcoRII may be applied in developing interventions against HSV-2 are presented: a nanoparticle for microbicide development and a "recombinant lactobacillus" expressing cell wall anchored receptor (truncated nectin-1) for HSV-2 plus EcoRII.

Conclusion: Viral genome slicing by way of these bacterially- derived R-M enzymatic peptides may have therapeutic potential in HSV-2 infection; a cofactor for HIV-1 acquisition and transmission.

Show MeSH
Related in: MedlinePlus