A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt)

Karine Nozeret (1, 2) , Alix Boucharlat (3) , Fabrice Agou (3) , Nienke Buddelmeijer (1, 2)

1 BGPB - Biologie et Génétique de la Paroi bactérienne - Biology and Genetics of Bacterial Cell Wall
2 Inserm Avenir Group
3 C2RT - Centre de Ressources et de Recherche Technologique - Center for Technological Resources and Research

Lipoprotein modification is an essential process in Gram-negative bacteria. The action of three integral membrane proteins that catalyze the transfer of fatty acids derived from membrane phospholipids or cleave the signal peptide of the lipoprotein substrate result in the formation of mature triacylated proteins. Inactivation of the enzymes leads to mis-localization of immature lipoproteins and consequently cell death. Biochemical studies and the development of in vitro assays are challenging due to the fact that the enzymes and substrates are all membrane-embedded proteins difficult to overproduce and purify. Here we describe a sensitive fluorescence-based assay to monitor bacterial apolipoprotein N-acyltransferase activity.

Domaines: Biochimie [q-bio.BM]; Bactériologie; Autre

Karine Nozeret, Alix Boucharlat, Fabrice Agou, Nienke Buddelmeijer. A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt). Scientific Reports, 2019, 9 (1), pp.15978. ⟨10.1038/s41598-019-52106-8⟩. ⟨pasteur-02421050⟩