Oxetane Grafts Installed Site‐Selectively on Native Disulfides to Enhance Protein Stability and Activity In Vivo
نویسندگان
چکیده
A four-membered oxygen ring (oxetane) can be readily grafted into native peptides and proteins through site-selective bis-alkylation of cysteine residues present as disulfides under mild and biocompatible conditions. The selective installation of the oxetane graft enhances stability and activity, as demonstrated for a range of biologically relevant cyclic peptides, including somatostatin, proteins, and antibodies, such as a Fab arm of the antibody Herceptin and a designed antibody DesAb-Aβ against the human Amyloid-β peptide. Oxetane grafting of the genetically detoxified diphtheria toxin CRM197 improves significantly the immunogenicity of this protein in mice, which illustrates the general utility of this strategy to modulate the stability and biological activity of therapeutic proteins containing disulfides in their structures.
منابع مشابه
Site – specific PEGylation of rHuEPO and evaluation of its biological activity and stability
Despite the critical role of erythropoietin (EPO) as therapeutic agent in treatment of anemia, its consumption is limited due to several disadvantages including the product short half-life, immunogenicity and susceptibility to proteolytic degradation. To overcome these drawbacks efficient methods such as site-specific PEGylation have been developed among witch N-terminal PEGylation has found mo...
متن کاملAntisense oligonuclotides with oxetane-constrained cytidine enhance heteroduplex stability, and elicit satisfactory RNase H response as well as showing improved resistance to both exo and endonucleases.
Antisense oligonucleotides (AONs) with single and double oxetane C modifications [1',2'-oxetane constrained cytidine, 1-(1',3'-O-anhydro-beta-D-psicofuranosyl)cytosine] have been evaluated, in comparison with the corresponding T-modified AONs, for their antisense potentials by targeting to a 15mer complementary RNA. Although the C modified mixmer AONs show approximately 3 degrees C drop per mod...
متن کاملStructural and Activity Comparison of Native, Apo and Reconstituted Tyrosinase
Background: Mushroom Tyrosinase a potent candidate in clinical studies known as polyphenol oxidase, is a metaloenzyme from the oxidase superfamily widely distributed from lower to higher life forms. It plays a crucial role in sclerotization of exoskeleton in insects, also responsible for skin pigmentation in mammalians. Objective: In this study, after reconstitution of MT by some metal io...
متن کاملOxetane modified antisense oligonucleotides promote RNase H cleavage of the complementary RNA strand in the hybrid duplex as efficiently as the native, and offer improved endonuclease resistance
Although the T m drops ∼6 C/modification (note: T m loss is ∼10 C/mismatch) in the oxetane, [1-(1 ,3 -Oanhydro-β--psicofuranosyl)thymine, T], modified antisense (AON)–RNA heteroduplexes, the relative rates of the complementary RNA cleavage by RNase H remain the same as or comparable to that of the native counterpart. The RNA cleavage in the native hybrid duplex was 68 ± 3% (T m = 44 C), wherea...
متن کاملStructural insights into the effects of charge-reversal substitutions at the surface of horseradish peroxidase
Horseradish peroxidase (HRP), has gained significant interests in biotechnology, especially in biosensor field and diagnostic test kits. Hence, its solvent-exposed lysine residues 174, 232, and 241 have been frequently modified with the aim of improving its stability and catalytic efficiency. In this computational study, we investigated the effects of Lys-to-Glu substitutions on HRP structure t...
متن کامل