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PDBsum entry 4y8h
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Hydrolase/hydrolase inhibitor
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PDB id
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4y8h
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Contents |
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250 a.a.
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244 a.a.
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240 a.a.
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235 a.a.
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231 a.a.
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243 a.a.
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241 a.a.
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222 a.a.
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204 a.a.
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195 a.a.
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212 a.a.
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222 a.a.
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233 a.a.
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196 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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Systematic analyses of substrate preferences of 20s proteasomes using peptidic epoxyketone inhibitors.
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Authors
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E.M.Huber,
G.De bruin,
W.Heinemeyer,
G.Paniagua soriano,
H.S.Overkleeft,
M.Groll.
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Ref.
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J Am Chem Soc, 2015,
137,
7835-7842.
[DOI no: ]
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PubMed id
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Abstract
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Cleavage analyses of 20S proteasomes with natural or synthetic substrates
allowed to infer the substrate specificities of the active sites and paved the
way for the rational design of high-affinity proteasome inhibitors. However,
details of cleavage preferences remained enigmatic due to the lack of
appropriate structural data. In a unique approach, we here systematically
examined substrate specificities of yeast and human proteasomes using
irreversibly acting α',β'epoxyketone (ep) inhibitors. Biochemical and
structural analyses provide unique insights into the substrate preferences of
the distinct active sites and highlight differences between proteasome types
that may be considered in future inhibitor design efforts. (1) For steric
reasons, epoxyketones with Val or Ile at the P1 position are weak inhibitors of
all active sites. (2) Identification of the β2c selective compound Ac-LAE-ep
represents a promising starting point for the development of compounds that
discriminate between β2c and β2i. (3) The compound Ac-LAA-ep was found to
favor subunit β5c over β5i by three orders of magnitude. (4) Yeast β1 and
human β1c subunits preferentially bind Asp and Leu in their S1 pockets, while
Glu and large hydrophobic residues are not accepted. (5) Exceptional structural
features in the β1/2 substrate binding channel give rise to the β1 selectivity
of compounds featuring Pro at the P3 site. Altogether, 23 different epoxyketone
inhibitors, five proteasome mutants, and 43 crystal structures served to
delineate a detailed picture of the substrate and ligand specificities of
proteasomes and will further guide drug development efforts toward
subunit-specific proteasome inhibitors for applications as diverse as cancer and
autoimmune disorders.
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