| Autor: |
Gahloth D; Manchester Institute of Biotechnology, University of Manchester, Manchester, UK., Fisher K; Manchester Institute of Biotechnology, University of Manchester, Manchester, UK., Marshall S; Manchester Institute of Biotechnology, University of Manchester, Manchester, UK., Leys D; Manchester Institute of Biotechnology, University of Manchester, Manchester, UK. Electronic address: david.leys@manchester.ac.uk. |
| Jazyk: |
angličtina |
| Zdroj: |
The Journal of biological chemistry [J Biol Chem] 2024 Feb; Vol. 300 (2), pp. 105653. Date of Electronic Publication: 2024 Jan 13. |
| DOI: |
10.1016/j.jbc.2024.105653 |
| Abstrakt: |
The UbiD enzyme family of prenylated flavin (prFMN)-dependent reversible decarboxylases is near ubiquitously present in microbes. For some UbiD family members, enzyme activation through prFMNH 2 binding and subsequent oxidative maturation of the cofactor readily occurs, both in vivo in a heterologous host and through in vitro reconstitution. However, isolation of the active holo-enzyme has proven intractable for others, notably the canonical Escherichia coli UbiD. We show that E. coli heterologous expression of the small protein LpdD-associated with the UbiD-like gallate decarboxylase LpdC from Lactobacillus plantarum-unexpectedly leads to 3,4-dihydroxybenzoic acid decarboxylation whole-cell activity. This activity was shown to be linked to endogenous E. coli ubiD expression levels. The crystal structure of the purified LpdD reveals a dimeric protein with structural similarity to the eukaryotic heterodimeric proteasome assembly chaperone Pba3/4. Solution studies demonstrate that LpdD protein specifically binds to reduced prFMN species only. The addition of the LpdD-prFMNH 2 complex supports reconstitution and activation of the purified E. coli apo-UbiD in vitro, leading to modest 3,4-dihydroxybenzoic acid decarboxylation. These observations suggest that LpdD acts as a prFMNH 2 -binding chaperone, enabling apo-UbiD activation through enhanced prFMNH 2 incorporation and subsequent oxidative maturation. Hence, while a single highly conserved flavin prenyltransferase UbiX is found associated with UbiD enzymes, our observations suggest considerable diversity in UbiD maturation, ranging from robust autocatalytic to chaperone-mediated processes. Unlocking the full (de)carboxylation scope of the UbiD-enzyme family will thus require more than UbiX coexpression.
Competing Interests: Conflict of interest The authors declare that they have no conflict of interest with the contents of this article.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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