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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.)

Structural Basis of the Selectivity of GenN, an Aminoglycoside N-Methyltransferase Involved in Gentamicin Biosynthesis

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Author(s):
Bury, Priscila dos Santos [1] ; Huang, Fanglu [2] ; Li, Sicong [3] ; Sun, Yuhui [3] ; Leadlay, Peter F. [2] ; Bertacine Dias, Marcio Vinicius [1]
Total Authors: 6
Affiliation:
[1] Univ Sao Paulo, Inst Biomed Sci, Dept Microbiol, Sao Paulo - Brazil
[2] Univ Cambridge, Dept Biochem, Cambridge CB2 1GA - England
[3] Wuhan Univ, Minist Educ, Sch Pharmaceut Sci, Key Lab Combinatorial Biosynth & Drug Discovery, Wuhan, Hubei - Peoples R China
Total Affiliations: 3
Document type: Journal article
Source: ACS Chemical Biology; v. 12, n. 11, p. 2779-2787, NOV 2017.
Web of Science Citations: 2
Abstract

Gentamicins are heavily methylated, clinically valuable pseudotrisaccharide antibiotics produced by Micromonospora echinospora. GenN has been characterized as an S-adenosyl-l-methionine-dependent methyltransferase with low sequence similarity to other enzymes. It is responsible for the 3?-N-methylation of 3 `'-dehydro-3 `'-amino-gentamicin A2, an essential modification of ring III in the biosynthetic pathway to the gentamicin C complex. Purified recombinant GenN also efficiently catalyzes 3 `'-N-methylation of related aminoglycosides kanamycin B and tobramycin, which both contain an additional hydroxymethyl group at the C5 `' position in ring III. We have obtained eight cocrystal structures of GenN, at a resolution of 2.2 angstrom or better, including the binary complex of GenN and S-adenosyl-l-homocysteine (SAH) and the ternary complexes of GenN, SAH, and several aminoglycosides. The GenN structure reveals several features not observed in any other N-methyltransferase that fit it for its role in gentamicin biosynthesis. These include a novel N-terminal domain that might be involved in protein:protein interaction with upstream enzymes of the gentamicin X2 biosynthesis and two long loops that are involved in aminoglycoside substrate recognition. In addition, the analysis of structures of GenN in complex with different ligands, supported by the results of active site mutagenesis, has allowed us to propose a catalytic mechanism and has revealed the structural basis for the surprising ability of native GenN to act on these alternative substrates. (AU)

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: 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: 14/50324-0 - Towards safer aminogrycoside antibiotics: structure, function and engineering studies of key biosynthetic enzymes
Grantee:Marcio Vinicius Bertacine Dias
Support Opportunities: Regular Research Grants