1ND4

Crystal structure of aminoglycoside-3'-phosphotransferase-IIa

Summary for 1ND4

• Entry DOI: 10.2210/pdb1nd4/pdb
• Descriptor: Aminoglycoside 3'-phosphotransferase, MAGNESIUM ION, ACETATE ION, ... (6 entities in total)
• Functional Keywords: phosphotransferase, protein kinase, atpase, kanamycin, transferase
• Biological source: Klebsiella pneumoniae
• Total number of polymer chains: 2
• Total formula weight: 59339.26

Authors

Nurizzo, D.,Shewry, S.C.,Baker, E.N.,Smith, C.A. (deposition date: 2002-12-06, release date: 2003-09-16, Last modification date: 2024-03-13)

Primary citation

Nurizzo, D.,Shewry, S.C.,Perlin, M.H.,Brown, S.A.,Dholakia, J.N.,Fuchs, R.L.,Deva, T.,Baker, E.N.,Smith, C.A.
The crystal structure of aminoglycoside-3'-phosphotransferase-IIa, an enzyme responsible for antibiotic resistance
J.Mol.Biol., 327:491-506, 2003

PubMed Abstract: A major factor in the emergence of antibiotic resistance is the existence of enzymes that chemically modify common antibiotics. The genes for these enzymes are commonly carried on mobile genetic elements, facilitating their spread. One such class of enzymes is the aminoglycoside phosphotransferase (APH) family, which uses ATP-mediated phosphate transfer to chemically modify and inactivate aminoglycoside antibiotics such as streptomycin and kanamycin. As part of a program to define the molecular basis for aminoglycoside recognition and inactivation by such enzymes, we have determined the high resolution (2.1A) crystal structure of aminoglycoside-3'-phosphotransferase-IIa (APH(3')-IIa) in complex with kanamycin. The structure was solved by molecular replacement using multiple models derived from the related aminoglycoside-3'-phosphotransferase-III enzyme (APH(3')-III), and refined to an R factor of 0.206 (R(free) 0.238). The bound kanamycin molecule is very well defined and occupies a highly negatively charged cleft formed by the C-terminal domain of the enzyme. Adjacent to this is the binding site for ATP, which can be modeled on the basis of nucleotide complexes of APH(3')-III; only one change is apparent with a loop, residues 28-34, in a position where it could fold over an incoming nucleotide. The three rings of the kanamycin occupy distinct sub-pockets in which a highly acidic loop, residues 151-166, and the C-terminal residues 260-264 play important parts in recognition. The A ring, the site of phosphoryl transfer, is adjacent to the catalytic base Asp190. These results give new information on the basis of aminoglycoside recognition, and on the relationship between this phosphotransferase family and the protein kinases.

PubMed: 12628253
DOI: 10.1016/S0022-2836(03)00121-9

Experimental method

X-RAY DIFFRACTION (2.1 Å)