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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
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]
Total Authors: 7
Affiliation:
[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
Total Affiliations: 4
Document type: Journal article
Source: ACS Chemical Biology; v. 14, n. 5, p. 925-933, MAY 2019.
Web of Science Citations: 0
Abstract

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)

FAPESP's process: 10/15971-3 - Structural characterization of enzymes from antibiotic biosynthetic pathways with biotechnological interest
Grantee:Marcio Vinicius Bertacine Dias
Support Opportunities: Research Grants - Young Investigators Grants
FAPESP's process: 15/09188-8 - Biosynthesis of polyether and aminoglycoside antibiotics: structural investigation of unusual enzymes or with synthetic biology applicability
Grantee:Marcio Vinicius Bertacine Dias
Support Opportunities: Regular Research Grants
FAPESP's process: 14/07843-6 - Structural analysis of key enzymes for gentamicin and sisomicin biosynthesis
Grantee:Priscila dos Santos Bury
Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 14/50324-0 - Towards safer aminogrycoside antibiotics: structure, function and engineering studies of key biosynthetic enzymes
Grantee:Marcio Vinicius Bertacine Dias
Support Opportunities: Regular Research Grants