 |
PDBsum entry 1oss
|
|
|
|
|
 |
Contents |
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
* Residue conservation analysis
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Engineering the primary substrate specificity of streptomyces griseus trypsin.
|
|
|
Authors
|
|
M.J.Page,
S.L.Wong,
J.Hewitt,
N.C.Strynadka,
R.T.Macgillivray.
|
|
|
Ref.
|
|
Biochemistry, 2003,
42,
9060-9066.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
Abstract
|
|
|
Streptomyces griseus trypsin (SGT) was chosen as a model scaffold for the
development of serine proteases with enhanced substrate specificity. Recombinant
SGT has been produced in a Bacillus subtilis expression system in a soluble
active form and purified to homogeneity. The recombinant and native proteases
have nearly identical enzymatic properties and structures. Four SGT mutants with
alterations in the S1 substrate binding pocket (T190A, T190P, T190S, and T190V)
were also expressed. The T190P mutant demonstrated the largest shift to a
preference for Arg versus Lys in the P1 site. This was shown by a minor
reduction in catalytic activity toward an Arg-containing substrate (k(cat)
reduction of 25%). The crystal structures of the recombinant SGT and the T190P
mutant in a complex with the inhibitor benzamidine were obtained at high
resolution (approximately 1.9 A). The increase in P1 specificity, achieved with
minimal effect on the catalytic efficiency, demonstrates that the T190P mutant
is an ideal candidate for the design of additional substrate specificity
engineered into the S2 to S4 binding pockets.
|
|
|
Secondary reference #1
|
|
|
Title
|
|
Refined crystal structure of streptomyces griseus trypsin at 1.7 a resolution.
|
|
|
Authors
|
|
R.J.Read,
M.N.James.
|
|
|
Ref.
|
|
J Mol Biol, 1988,
200,
523-551.
|
|
|
PubMed id
|
|
|
|
|
|
|
|
|
Secondary reference #2
|
|
|
Title
|
|
Critical evaluation of comparative model building of streptomyces griseus trypsin.
|
|
|
Authors
|
|
R.J.Read,
G.D.Brayer,
L.Jurásek,
M.N.James.
|
|
|
Ref.
|
|
Biochemistry, 1984,
23,
6570-6575.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
|
|
|
|
|
|
|
