5NCI

GriE in complex with cobalt, alpha-ketoglutarate and l-leucine

Summary for 5NCI

• Entry DOI: 10.2210/pdb5nci/pdb
• Descriptor: Leucine hydroxylase, COBALT (II) ION, 2-OXOGLUTARIC ACID, ... (6 entities in total)
• Functional Keywords: fe(ii)/alpha-ketoglutarate dependent dioxygenase, hydroxylase, non-heme iron protein, leucine hydroxylase, 5-hydroxyleucine, (2s4r)-5-hydroxyleucine, 4-methyl-proline, griselimycin, methyl-griselimycin, oxidoreductase
• Biological source: Streptomyces sp. DSM 40835
• Total number of polymer chains: 2
• Total formula weight: 60424.38

Authors

Lukat, P.,Blankenfeldt, W.,Mueller, R. (deposition date: 2017-03-05, release date: 2017-10-04, Last modification date: 2024-01-17)

Primary citation

Lukat, P.,Katsuyama, Y.,Wenzel, S.,Binz, T.,Konig, C.,Blankenfeldt, W.,Bronstrup, M.,Muller, R.
Biosynthesis of methyl-proline containing griselimycins, natural products with anti-tuberculosis activity.
Chem Sci, 8:7521-7527, 2017

PubMed Abstract: Griselimycins (GMs) are depsidecapeptides with superb anti-tuberculosis activity. They contain up to three (2,4)-4-methyl-prolines (4-MePro), of which one blocks oxidative degradation and increases metabolic stability in animal models. The natural congener with this substitution is only a minor component in fermentation cultures. We showed that this product can be significantly increased by feeding the reaction with 4-MePro and we investigated the molecular basis of 4-MePro biosynthesis and incorporation. We identified the GM biosynthetic gene cluster as encoding a nonribosomal peptide synthetase and a sub-operon for 4-MePro formation. Using heterologous expression, gene inactivation, and experiments, we showed that 4-MePro is generated by leucine hydroxylation, oxidation to an aldehyde, and ring closure with subsequent reduction. The crystal structures of the leucine hydroxylase GriE have been determined in complex with substrates and products, providing insight into the stereospecificity of the reaction.

PubMed: 29163906
DOI: 10.1039/c7sc02622f

Experimental method

X-RAY DIFFRACTION (1.755 Å)