Structure of IDP91191

Crystal structure of the aminoglycoside phosphotransferase APH(3')-Ia, with substrate kanamycin and small molecule inhibitor anthrapyrazolone SP600125

Edit deposit information
CSGID target
IDP91191 
PDB Id
4FEU (NCBI MMDB
Authors
P.J.Stogios,E.Evdokimova,Z.Wawrzak,G.Minasov,O.Egorova,R.Di Leo,T.Shakya,P.Spanogiannopoulos,G.D.Wright,A.Savchenko,W.F.Anderson,Center For Structural Genomics Of Infectious Diseases (Csgid) 
Responsible person
Peter Stogios 
Responsible lab
University of Calgary 
Deposition Date
May 30, 2012 
Release Date
Jun 20, 2012 

Annotation

Description
Activity of the aminoglycoside phosphotransferase APH(3')-Ia leads to resistance to aminoglycoside antibiotics in pathogenic Gram-negative bacteria, and contributes to the clinical obsolescence of this class of antibiotics. One strategy to rescuing compromised antibiotics such as aminoglycosides is targeting the enzymes conferring resistance with small molecules. Previously we demonstrated that eukaryotic protein kinase (ePK) inhibitors could inhibit APH enzymes, due to the structural similarity between these two enzyme families. However, limited structural information of enzyme-inhibitor complexes hindered interpretation of the results. As well, cross-reactivity of compounds between APHs and ePKs represents an obstacle to their use as aminoglycoside adjuvants to rescue aminoglycoside antibiotic activity. Here, we structurally and functionally characterize inhibition of APH(3')-Ia by three diverse chemical scaffolds - anthrapyrazolone, 4-anilinoquinazoline and pyrazolopyrimidine (PP) - and reveal distinctions in the binding mode of anthrapyrazolone and PP compounds to APH(3')-Ia versus ePKs. Using this observation, we identify PP-derivatives that select against ePKs, attenuate APH(3')-Ia activity and rescue aminoglycoside antibiotic activity against a resistant E. coli strain. The structures presented here and these inhibition studies provide an important opportunity for structure-based design of compounds to target aminoglycoside phosphotransferases for inhibition, potentially overcoming this form of antibiotic resistance.  
Functional assignment
antibiotic resistance 

Ligands

Ligand code Name Ligand type
KAN crystallization
537 2,6-dihydroanthra/1,9-cd/pyrazol-6-one crystallization
ACT acetate crystallization
175 3,5-dihydro-5-methylidene-4h-imidazol-4-on

Structure information

Unit cell parameters

Space Group
P 1  
Unit Cell

a=57.75Å, b=94.16Å, c=96.77Å
α=61.21, β=73.11, γ=87.36 
Solvent content
 
Matthews coefficient
 

Refinement

Data for the highest resolution shell is in parentheses.
Resolution range
19.97-2.37Å (0.00-0.00Å)  
Rall(%)
16.3 
Rwork(%)
16.0 (18.8) 
Rfree(%)
21.5 (21.8) 
Num. observed reflections
70887 (4752) 
Num. Rfree reflections
3579 (218) 
Completeness(%)
0.0 (0.0) 

Model parameters

Num Atoms
12085  
Num Waters
675  
Num Hetatoms
1246  
Model mean isotropic B factor
55.880Å2  
RMSD bond length
0.010Å  
RMSD bond angle
2.800°  
RMSD improper torsion angle
18.48°
 
Filename uploaded
4FEU.pdb (uploaded on Jun 26, 2013 10:40 AM)  
Inserted
May 30, 2012