Crystal Structure of GenD2, an NAD-Dependent Oxidoreductase Involved in the Biosynthesis of Gentamicin

Autor(es)

de Araujo, Natalia Cerrone ^[1] ; Bury, Priscila dos Santos ^[1] ; Tavares, Mauricio Temotheo ^[2] ; Huang, Fanglu ^[3] ; Parise-Filho, Roberto ^[2] ; Leadlay, Peter ^[3] ; Bertacine Dias, Marcio Vinicius ^{[4, 1]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Biomed Sci, Dept Microbiol, Ave Prof Lineu Prestes 1374, BR-05508900 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Fac Pharmaceut Sci, Dept Pharm, Prof Lineu Prestes Ave 580, BR-05508900 Sao Paulo - Brazil

^[3] Univ Cambridge, Dept Biochem, 80 Tennis Court Rd, Cambridge CB2 1GA - England

^[4] Univ Warwick, Dept Chem, Coventry CV4 7AL, W Midlands - England

• Tipo de documento: Artigo Científico
• Fonte: ACS Chemical Biology; v. 14, n. 5, p. 925-933, MAY 2019.
• Citações Web of Science: 0

Resumo

Gentamicins are clinically relevant aminoglycoside antibiotics produced by several Micromonospora species. Gentamicins are highly methylated and functionalized molecules, and their biosynthesis include glycosyltransferases, dehydratase/oxidoreductases, aminotransferases, and methyltransferases. The biosynthesis of gentamicin A from gentamicin A2 involves three enzymatic steps that modify the hydroxyl group at position 3 `' of the unusual garosamine sugar to provide its substitution for an amino group, followed by an N-methylation. The first of these reactions is catalyzed by GenD2, an oxidoreductase from the Gfo/Idh/MocA protein family, which reduces the hydroxyl at the C3 `' of gentamicin A to produce 3 `'-dehydro-3 `'-oxo-gentamicin A2 (DOA2). In this work, we solved the structure of GenD2 in complex with NAD+. Although the structure of GenD2 has a similar fold to other members of the Gfo/Idh/MocA family, this enzyme has several new features, including a 3D-domain swapping of two beta-strands that are involved in a novel oligomerization interface for this protein family. In addition, the active site of this enzyme also has several specialties which are possibly involved in the substrate specificity, including a number of aromatic residues and a negatively charged region, which is complementary to the polycationic aminoglycoside-substrate. Therefore, docking simulations provided insights into the recognition of gentamicin A2 and into the catalytic mechanism of GenD2. This is the first report describing the structure of an oxidoreductase involved in aminoglycoside biosynthesis and could open perspectives into producing new aminoglycoside derivatives by protein engineering. (AU)

• Processo FAPESP: 10/15971-3 - Caracterização estrutural de enzimas envolvidas em vias de biossíntese de antibióticos com interesse biotecnológico
• Beneficiário: Marcio Vinicius Bertacine Dias
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 15/09188-8 - Biossíntese de antibióticos poliéteres e aminoglicosídeos: investigação estrutural de enzimas não usuais ou com aplicabilidade em biologia sintética
• Beneficiário: Marcio Vinicius Bertacine Dias
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/07843-6 - Análise estrutural de enzimas chaves para biossíntese de gentamicina e sisomicina
• Beneficiário: Priscila dos Santos Bury
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 14/50324-0 - Produção de antibióticos aminoglicosídeos menos tóxicos: estrutura, função e estudo de bioengenharia de enzimas biossintéticas chaves
• Beneficiário: Marcio Vinicius Bertacine Dias
• Modalidade de apoio: Auxílio à Pesquisa - Regular