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

Kód výsledku v 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>

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

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

Rok uplatnění

2017

Kód důvěrnosti údajů

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

Název periodika

FEBS Open Bio

ISSN

2211-5463

e-ISSN

—

Svazek periodika

7

Číslo periodika v rámci svazku

12

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

16

Strana od-do

1864-1879

Kód UT WoS článku

000417109000004

EID výsledku v databázi Scopus

2-s2.0-85037166760