The Radical SAM enzyme NirJ catalyzes the removal of two propionate side chains during hemed1biosynthesis
| Autor: | Linda Boss, Susan Billig, Claudia Birkemeyer, Gunhild Layer, Ramona Oehme |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification biology Stereochemistry Substrate (chemistry) Cell Biology Biochemistry Tetrapyrrole Cofactor Enzyme assay 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology chemistry biology.protein Propionate Molecular Biology Radical SAM Heme Pyrrole |
| Zdroj: | The FEBS Journal. 284:4314-4327 |
| ISSN: | 1742-464X |
| DOI: | 10.1111/febs.14307 |
| Popis: | Heme d1 is a modified tetrapyrrole playing an important role in denitrification by acting as the catalytically essential cofactor in the cytochrome cd1 nitrite reductase of many denitrifying bacteria. In the course of heme d1 biosynthesis, the two propionate side chains on pyrrole rings A and B of the intermediate 12,18-didecarboxysiroheme are removed from the tetrapyrrole macrocycle. In the final heme d1 molecule, the propionate groups are replaced by two keto functions. Although it was speculated that the Radical S-adenosyl-l-methionine (SAM) enzyme NirJ might be responsible for the removal of the propionate groups and introduction of the keto functions, this has not been shown experimentally, so far. Here, we demonstrate that NirJ is a Radical SAM enzyme carrying two iron-sulfur clusters. While the N-terminal [4Fe-4S] cluster is essential for the initial SAM cleavage reaction, it is not required for substrate binding. NirJ tightly binds its substrate 12,18-didecarboxysiroheme and, thus, can be purified in complex with the substrate. By using the purified NirJ/substrate complex in an in vitro enzyme activity assay, we show that NirJ indeed catalyzes the removal of the two propionate side chains under simultaneous SAM cleavage. However, under the reaction conditions employed, no keto group formation is observed indicating that an additional cofactor or enzyme is needed for this reaction. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|