ABSTRACT
HPV-2 is a very common type of HPV which causes common warts. The E2 protein of virus can repress the activity of the viral early promoter through binding to the specific binding sites in viral LCR. Previously we reported that the repression of a mutated E2 protein of HPV-2 isolated from a patient with huge common wart on the viral early promoter was obviously decreased, and A338V mutation located at the C terminal DNA binding region of E2 protein. In this study, we expressed and purified the recombinant mutated and prototype E2 fusion proteins, both in the contexts of the C terminal and the full length, by prokaryotic expression system. The electrophoretic mobility shift assay showed E2 protein could bind to double-stranded DNA oligos labeled with biotin that covered two E2 binding sites. The DNA binding abilities of both C terminal and full-length mutated E2 proteins were stronger than the prototype analogs. This result indicates that the enhancement of the mutated E2 DNA binding ability may be the molecular mechanism for its impact on the activity of viral promoter, which correlates with the phenotype of extensive common wart.
Subject(s)
DNA , Metabolism , DNA-Binding Proteins , Genetics , Metabolism , Electrophoresis , Genetic Vectors , Genetics , Mutation , Papillomaviridae , Promoter Regions, Genetic , Genetics , Protein Binding , Recombinant Proteins , Genetics , Metabolism , Viral Proteins , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To develop and investigate GLL-37, a substitution analogue of the human antimicrobial peptide LL-37 with anti-enzymatic degradation activity and improved efficacy.</p><p><b>METHODS</b>The bactericidal activities of LL-37 and newly developed GLL-37 against 6 Gram-negative and -positive bacteria were determined by Broth microdilution assays. The minimum inhibitory concentrations of LL-37 and GLL-37 against E.coli ATCC 25922 in different NaCl concentration medium were also detected. Both peptides were co-incubated with elastase, and then analyzed by PAGE electrophoresis and bactericidal activity determination.</p><p><b>RESULT</b>GLL-37 showed a stronger elastase resistance ability than LL-37, and was significantly more effective than LL-37 under high-salt condition.</p><p><b>CONCLUSION</b>The antimicrobial peptide GLL-37 derived form LL-37 has the potential as a new therapeutic agent for bacterial infections.</p>
Subject(s)
Animals , Female , Humans , Anti-Bacterial Agents , Pharmacology , Therapeutic Uses , Antimicrobial Cationic Peptides , Pharmacology , Therapeutic Uses , Blood Bactericidal Activity , Cathelicidins , Cell Membrane Permeability , Escherichia coli , Membrane Proteins , Metabolism , Monocytes , Pseudomonas Infections , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To investigate the analgesic effect of interleukin-2 (IL-2) in mice with spared nerve injury (SNI).</p><p><b>METHOD</b>IL-2 was intraperitoneally injected in mice with induced SNI, and von Frey Filaments test and cold plate test were carried out to accesses the analgesic effects of IL-2 and the effect of naloxone in antagonizing the effects of IL-2.</p><p><b>RESULTS</b>IL-2 produced analgesic effects against hyperalgesia and allodynia in mouse models of SNI, and the effect of IL-2 lasted for more than 24 h, showing a double-peak pattern in its action with the two peaks occurring at 30 and 105 min, respectively. The effect of IL-2 could be significantly antagonized by naloxone.</p><p><b>CONCLUSIONS</b>IL-2 has long-lasting analgesic effects in mouse models of SNI model, showing a double-peak pattern of its action. The analgesic effect of IL-2 is probably mediated by opiate receptor.</p>