Naslov
Modeling and experimental analyses reveal a two-domain structure and amino acids important for the activity of aminoglycoside resistance methyltransferase Sgm

Autori
Maravić Vlahoviček, Gordana ; Čubrilo, Sonja ; Tkaczuk, Karolina L. ; Bujnicki, Janusz M.

Izvornik
Biochimica et Biophysica Acta - Proteins and Proteomics (1570-9639) 1784 (2008), 4; 582-90

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
Antibiotic resistance; Methylation; Modeling; Site-directed mutagenesis

Sažetak
Methyltransferases that carry out posttranscriptional N7-methylation of G1405 in 16S rRNA confer bacterial resistance to aminoglycoside antibiotics, including kanamycin and gentamicin. Genes encoding enzymes from this family (hereafter referred to as Arm, for aminoglycoside resistance methyltransferases) have been recently found to spread by horizontal gene transfer between various human pathogens. The knowledge of the Arm protein structure would lay the groundwork for the development of potential resistance inhibitors, which could be used to restore the potential of aminoglycosides to act against the resistant pathogens. We analyzed the sequence– function relationships of Sgm MTase, a member of the Arm family, by limited proteolysis and site-directed and random mutagenesis. We also modeled the structure of Sgm using bioinformatics techniques and used the model to provide a structural context for experimental results. We found that Sgm comprises two domains and we characterized a number of functionally compromised point mutants with substitutions of invariant or conserved residues. Our study provides a low-resolution (residue-level) model of sequence– structure– function relationships in the Arm family of enzymes and reveals the cofactor-binding and substrate-binding sites. These functional regions will be prime targets for further experimental and theoretical studies aimed at defining the reaction mechanism of m7G1405 methylation, increasing the resolution of the model and developing Arm-specific inhibitors.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA