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PDBsum entry 2bkk
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Transferase/peptide
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PDB id
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2bkk
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Contents |
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247 a.a.
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156 a.a.
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262 a.a.
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References listed in PDB file
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Key reference
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Title
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Allosteric inhibition of aminoglycoside phosphotransferase by a designed ankyrin repeat protein.
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Authors
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A.Kohl,
P.Amstutz,
P.Parizek,
H.K.Binz,
C.Briand,
G.Capitani,
P.Forrer,
A.Plückthun,
M.G.Grütter.
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Ref.
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Structure (Camb), 2005,
13,
1131-1141.
[DOI no: ]
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PubMed id
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Abstract
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Aminoglycoside phosphotransferase (3')-IIIa (APH) is a bacterial kinase that
confers antibiotic resistance to many pathogenic bacteria and shares structural
homology with eukaryotic protein kinases. We report here the crystal structure
of APH, trapped in an inactive conformation by a tailor-made inhibitory ankyrin
repeat (AR) protein, at 2.15 A resolution. The inhibitor was selected from a
combinatorial library of designed AR proteins. The AR protein binds the
C-terminal lobe of APH and thereby stabilizes three alpha helices, which are
necessary for substrate binding, in a significantly displaced conformation.
BIAcore analysis and kinetic enzyme inhibition experiments are consistent with
the proposed allosteric inhibition mechanism. In contrast to most small-molecule
kinase inhibitors, the AR proteins are not restricted to active site binding,
allowing for higher specificity. Inactive conformations of pharmaceutically
relevant enzymes, as can be elucidated with the approach presented here,
represent powerful starting points for rational drug design.
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Figure 2.
Figure 2. Crystal Structure of the mAPH in Complex with the
AR Protein AR_3a
(A) Stereo view of the AB heterodimer of
the mAPH/AR_3a complex. The mAPH is shown in orange, and the
AR_3a is shown in light blue. AR_3a binds to the C-terminal lobe
of the mAPH and stabilizes an inactive conformation.
(B)
Asymmetric mAPH pseudo-homodimer found in the crystal of the
mAPH/AR_3a complex.
(C) Symmetric wtAPH dimer found in the
wtAPH crystal.
(D) Stereo view of the superposition of the
mAPH hetereodimer AB (orange) and BC (light blue).
(E)
wtAPH in the kanamycin A bound form (PDB: 1L8T1L8T). ADP and
kanamycin A are labeled.
(F) mAPH/AR_3a AB heterodimer in
the same orientation as the wtAPH in (E).
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The above figure is
reprinted
by permission from Cell Press:
Structure (Camb)
(2005,
13,
1131-1141)
copyright 2005.
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