 |
PDBsum entry 1o5e
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Serine protease, hydrolase
|
PDB id
|
|
|
|
1o5e
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
Contents |
 |
|
|
|
|
|
|
|
|
|
|
|
|
* Residue conservation analysis
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Dissecting and designing inhibitor selectivity determinants at the s1 site using an artificial ala190 protease (ala190 upa).
|
|
|
Authors
|
|
B.A.Katz,
C.Luong,
J.D.Ho,
J.R.Somoza,
E.Gjerstad,
J.Tang,
S.R.Williams,
E.Verner,
R.L.Mackman,
W.B.Young,
P.A.Sprengeler,
H.Chan,
K.Mortara,
J.W.Janc,
M.E.Mcgrath.
|
|
|
Ref.
|
|
J Mol Biol, 2004,
344,
527-547.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
Abstract
|
|
|
A site-directed mutant of the serine protease urokinase-type plasminogen
activator (uPA), was produced to assess the contribution of the Ser190
side-chain to the affinity and selectivity of lead uPA inhibitors in the absence
of other differences present in comparisons of natural proteases.
Crystallography and enzymology involving WT and Ala190 uPA were used to
calculate free energy binding contributions of hydrogen bonds involving the
Ser190 hydroxyl group (O(gamma)(Ser190)) responsible for the remarkable
selectivity of 6-halo-5-amidinoindole and 6-halo-5-amidinobenzimidazole
inhibitors toward uPA and against natural Ala190 protease anti-targets. Crystal
structures of uPA complexes of novel, active site-directed arylguanidine and
2-aminobenzimidazole inhibitors of WT uPA, together with associated K(i) values
for WT and Ala190 uPA, also indicate a significant role of Ser190 in the binding
of these classes of uPA inhibitors. Structures and associated K(i) values for a
lead inhibitor (CA-11) bound to uPA and to five other proteases, as well as for
other leads bound to multiple proteases, help reveal the features responsible
for the potency (K(i)=11nM) and selectivity of the remarkably small inhibitor,
CA-11. The 6-fluoro-5-amidinobenzimidzole, CA-11, is more than 1000-fold
selective against natural Ala190 protease anti-targets, and more than 100-fold
selective against other Ser190 anti-targets.
|
|
|
|
|
|
|
|
Figure 2.
Figure 2. Structure and associated (|F[o]| -|F[c]|), a[c]
omit map for hepsin-CA-14, contoured at 2.5s.
|
|
Figure 9.
Figure 9. S1 site structure of the complex of WT uPA with
the (a) 6-fluoro (CA-10) inhibitor and (b) the non-fluoro analog
(CA-06). Short hydrogen bonds at the active site are cyan.
|
|
|
|
|
|
The above figures are
reprinted
by permission from Elsevier:
J Mol Biol
(2004,
344,
527-547)
copyright 2004.
|
|
|
|
|
|
|
