Characterization of auxiliary iron-sulfur clusters in a radical S-adenosylmethionine enzyme PqqE from Methylobacterium extorquens AM1

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F17%3A73584728" target="_blank" >RIV/61989592:15310/17:73584728 - isvavai.cz</a>

Result on the web

<a href="http://onlinelibrary.wiley.com/doi/10.1002/2211-5463.12314/epdf" target="_blank" >http://onlinelibrary.wiley.com/doi/10.1002/2211-5463.12314/epdf</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/2211-5463.12314" target="_blank" >10.1002/2211-5463.12314</a>

Result language

angličtina

Original language name

Characterization of auxiliary iron-sulfur clusters in a radical S-adenosylmethionine enzyme PqqE from Methylobacterium extorquens AM1

Original language description

PqqE is a radical S-adenosyl-l-methionine (SAM) enzyme that catalyzes the initial reaction of pyrroloquinoline quinone (PQQ) biosynthesis. PqqE belongs to the SPASM (subtilosin/PQQ/anaerobic sulfatase/mycofactocin maturating enzymes) subfamily of the radical SAM superfamily and contains multiple Fe-S clusters. To characterize the Fe-S clusters in PqqE from Methylobacterium extorquens AM1, Cys residues conserved in the N-terminal signature motif (CX3CX2C) and the C-terminal seven-cysteine motif (CX(9-15)GX(4)CX(n)CX(2)CX(5)CX(3)CX(n)C; n=an unspecified number) were individually or simultaneously mutated into Ser. Biochemical and Mossbauer spectral analyses of as-purified and reconstituted mutant enzymes confirmed the presence of three Fe-S clusters in PqqE: one [4Fe-4S](2+) cluster at the N-terminal region that is essential for the reductive homolytic cleavage of SAM into methionine and 5-deoxyadenosyl radical, and one each [4Fe-4S](2+) and [2Fe-2S](2+) auxiliary clusters in the C-terminal SPASM domain, which are assumed to serve for electron transfer between the buried active site and the protein surface. The presence of [2Fe-2S](2+) cluster is a novel finding for radical SAM enzyme belonging to the SPASM subfamily. Moreover, we found uncommon ligation of the auxiliary [4Fe-4S](2+) cluster with sulfur atoms of three Cys residues and a carboxyl oxygen atom of a conserved Asp residue.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

FEBS Open Bio

ISSN

2211-5463

e-ISSN

—

Volume of the periodical

7

Issue of the periodical within the volume

12

Country of publishing house

GB - UNITED KINGDOM

Number of pages

16

Pages from-to

1864-1879

UT code for WoS article

000417109000004

EID of the result in the Scopus database

2-s2.0-85037166760