Biochemical and structural characterisation of the second oxidative crosslinking step during the biosynthesis of the glycopeptide antibiotic A47934
| Autor: | Clara Brieke, Max J. Cryle, Veronika Ulrich |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
crystal structure cytochrome P450 medicine.drug_class Peptide Glycopeptide antibiotic Full Research Paper lcsh:QD241-441 03 medical and health sciences chemistry.chemical_compound phenolic coupling lcsh:Organic chemistry Biosynthesis medicine glycopeptide antibiotic lcsh:Science chemistry.chemical_classification biology Organic Chemistry Cytochrome P450 Substrate (chemistry) Biological activity peptide Glycopeptide Chemistry 030104 developmental biology Enzyme Biochemistry chemistry biology.protein lcsh:Q |
| Zdroj: | Beilstein Journal of Organic Chemistry, Vol 12, Iss 1, Pp 2849-2864 (2016) Beilstein Journal of Organic Chemistry |
| ISSN: | 1860-5397 |
| Popis: | The chemical complexity and biological activity of the glycopeptide antibiotics (GPAs) stems from their unique crosslinked structure, which is generated by the actions of cytochrome P450 (Oxy) enzymes that affect the crosslinking of aromatic side chains of amino acid residues contained within the GPA heptapeptide precursor. Given the crucial role peptide cyclisation plays in GPA activity, the characterisation of this process is of great importance in understanding the biosynthesis of these important antibiotics. Here, we report the cyclisation activity and crystal structure of StaF, the D-O-E ring forming Oxy enzyme from A47934 biosynthesis. Our results show that the specificity of StaF is reduced when compared to Oxy enzymes catalysing C-O-D ring formation and that this activity relies on interactions with the non-ribosomal peptide synthetase via the X-domain. Despite the interaction of StaF with the A47934 X-domain being weaker than for the preceding Oxy enzyme StaH, StaF retains higher levels of in vitro activity: we postulate that this is due to the ability of the StaF/X-domain complex to allow substrate reorganisation after initial complex formation has occurred. These results highlight the importance of testing different peptide/protein carrier constructs for in vitro GPA cyclisation assays and show that different Oxy homologues can display significantly different catalytic propensities despite their overall similarities. |
| Databáze: | OpenAIRE |
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