1os8: Difference between revisions
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[[Image:1os8.jpg|left|200px]] | [[Image:1os8.jpg|left|200px]] | ||
'''RECOMBINANT STREPTOMYCES GRISEUS TRYPSIN''' | |||
{{Structure | |||
|PDB= 1os8 |SIZE=350|CAPTION= <scene name='initialview01'>1os8</scene>, resolution 1.55Å | |||
|SITE= | |||
|LIGAND= <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene> and <scene name='pdbligand=CA:CALCIUM ION'>CA</scene> | |||
|ACTIVITY= [http://en.wikipedia.org/wiki/Trypsin Trypsin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.4 3.4.21.4] | |||
|GENE= SPRT ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1911 Streptomyces griseus]) | |||
}} | |||
'''RECOMBINANT STREPTOMYCES GRISEUS TRYPSIN''' | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
1OS8 is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_griseus Streptomyces griseus | 1OS8 is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_griseus Streptomyces griseus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1OS8 OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: trypsin]] | [[Category: trypsin]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 13:14:56 2008'' | ||
Revision as of 14:14, 20 March 2008
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| , resolution 1.55Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | and | ||||||
| Gene: | SPRT (Streptomyces griseus) | ||||||
| Activity: | Trypsin, with EC number 3.4.21.4 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
RECOMBINANT STREPTOMYCES GRISEUS TRYPSIN
OverviewOverview
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.
About this StructureAbout this Structure
1OS8 is a Single protein structure of sequence from Streptomyces griseus. Full crystallographic information is available from OCA.
ReferenceReference
Engineering the primary substrate specificity of Streptomyces griseus trypsin., Page MJ, Wong SL, Hewitt J, Strynadka NC, MacGillivray RT, Biochemistry. 2003 Aug 5;42(30):9060-6. PMID:12885239
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