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PDBsum entry 4cth
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References listed in PDB file
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Key reference
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Title
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Engineering neprilysin activity and specificity to create a novel therapeutic for alzheimer'S disease.
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Authors
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C.I.Webster,
M.Burrell,
L.L.Olsson,
S.B.Fowler,
S.Digby,
A.Sandercock,
A.Snijder,
J.Tebbe,
U.Haupts,
J.Grudzinska,
L.Jermutus,
C.Andersson.
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Ref.
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Plos One, 2014,
9,
e104001.
[DOI no: ]
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PubMed id
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Abstract
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Neprilysin is a transmembrane zinc metallopeptidase that degrades a wide range
of peptide substrates. It has received attention as a potential therapy for
Alzheimer's disease due to its ability to degrade the peptide amyloid beta.
However, its broad range of peptide substrates has the potential to limit its
therapeutic use due to degradation of additional peptides substrates that
tightly regulate many physiological processes. We sought to generate a soluble
version of the ectodomain of neprilysin with improved activity and specificity
towards amyloid beta as a potential therapeutic for Alzheimer's disease.
Extensive amino acid substitutions were performed at positions surrounding the
active site and inner surface of the enzyme and variants screened for activity
on amyloid beta 1-40, 1-42 and a variety of other physiologically relevant
peptides. We identified several mutations that modulated and improved both
enzyme selectivity and intrinsic activity. Neprilysin variant G399V/G714K
displayed an approximately 20-fold improved activity on amyloid beta 1-40 and up
to a 3,200-fold reduction in activity on other peptides. Along with the altered
peptide substrate specificity, the mutant enzyme produced a markedly altered
series of amyloid beta cleavage products compared to the wild-type enzyme.
Crystallisation of the mutant enzyme revealed that the amino acid substitutions
result in alteration of the shape and size of the pocket containing the active
site compared to the wild-type enzyme. The mutant enzyme offers the potential
for the more efficient degradation of amyloid beta in vivo as a therapeutic for
the treatment of Alzheimer's disease.
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