1f4d: Difference between revisions

Revision as of 15:53, 2 May 2008

CRYSTAL STRUCTURE OF E. COLI THYMIDYLATE SYNTHASE C146S, L143C COVALENTLY MODIFIED AT C143 WITH N-[TOSYL-D-PROLINYL]AMINO-ETHANETHIOL

OverviewOverview

We report a strategy (called "tethering") to discover low molecular weight ligands ( approximately 250 Da) that bind weakly to targeted sites on proteins through an intermediary disulfide tether. A native or engineered cysteine in a protein is allowed to react reversibly with a small library of disulfide-containing molecules ( approximately 1,200 compounds) at concentrations typically used in drug screening (10 to 200 microM). The cysteine-captured ligands, which are readily identified by MS, are among the most stable complexes, even though in the absence of the covalent tether the ligands may bind very weakly. This method was applied to generate a potent inhibitor for thymidylate synthase, an essential enzyme in pyrimidine metabolism with therapeutic applications in cancer and infectious diseases. The affinity of the untethered ligand (K(i) approximately 1 mM) was improved 3,000-fold by synthesis of a small set of analogs with the aid of crystallographic structures of the tethered complex. Such site-directed ligand discovery allows one to nucleate drug design from a spatially targeted lead fragment.

About this StructureAbout this Structure

1F4D is a Single protein structure of sequence from Escherichia coli. Full crystallographic information is available from OCA.

ReferenceReference

Site-directed ligand discovery., Erlanson DA, Braisted AC, Raphael DR, Randal M, Stroud RM, Gordon EM, Wells JA, Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9367-72. PMID:10944209 Page seeded by OCA on Fri May 2 15:52:59 2008

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

@@ Line 1: / Line 1: @@
 [[Image:1f4d.jpg|left|200px]]
- {{Structure
+<!--
-|PDB= 1f4d |SIZE=350|CAPTION= <scene name='initialview01'>1f4d</scene>, resolution 2.15&Aring;
+The line below this paragraph, containing "STRUCTURE_1f4d", creates the "Structure Box" on the page.
-|SITE=
+You may change the PDB parameter (which sets the PDB file loaded into the applet)
-|LIGAND= <scene name='pdbligand=CXM:N-CARBOXYMETHIONINE'>CXM</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>, <scene name='pdbligand=TP2:N-[TOSYL-D-PROLINYL]AMINO-ETHANETHIOL'>TP2</scene>
+or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
-|ACTIVITY= <span class='plainlinks'>[http://en.wikipedia.org/wiki/Thymidylate_synthase Thymidylate synthase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.1.1.45 2.1.1.45] </span>
+or leave the SCENE parameter empty for the default display.
-|GENE=
+-->
-|DOMAIN=
+{{STRUCTURE_1f4d|  PDB=1f4d  |  SCENE=  }}
-|RELATEDENTRY=[[1f4b|1F4B]], [[1f4c|1F4C]], [[1f4e|1F4E]], [[1f4f|1F4F]], [[1f4g|1F4G]]
-|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1f4d FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1f4d OCA], [http://www.ebi.ac.uk/pdbsum/1f4d PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1f4d RCSB]</span>
-}}
 '''CRYSTAL STRUCTURE OF E. COLI THYMIDYLATE SYNTHASE C146S, L143C COVALENTLY MODIFIED AT C143 WITH N-[TOSYL-D-PROLINYL]AMINO-ETHANETHIOL'''
@@ Line 33: / Line 30: @@
 [[Category: Stroud, R M.]]
 [[Category: Wells, J A.]]
-[[Category: crystal structure of e. coli thymidylate synthase c146]]
+[[Category: Crystal structure of e. coli thymidylate synthase c146]]
-[[Category: l143c covalently modified at c143 with n-[tosyl-d-prolinyl]amino-ethanethiol]]
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May  2 15:52:59 2008''
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 20:15:35 2008''