1f4c: Difference between revisions

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[[Image:1f4c.jpg|left|200px]]<br /><applet load="1f4c" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:1f4c.jpg|left|200px]]<br /><applet load="1f4c" size="350" color="white" frame="true" align="right" spinBox="true"

caption="1f4c, resolution 2.0Å" />

'''CRYSTAL STRUCTURE OF E. COLI THYMIDYLATE SYNTHASE COVALENTLY MODIFIED AT C146 WITH N-[TOSYL-D-PROLINYL]AMINO-ETHANETHIOL'''<br />

==Overview==

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.

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 Structure==

1F4C is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with SO4, TP2 and GOL as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [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] Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=1F4C OCA].

1F4C is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with <scene name='pdbligand=SO4:'>SO4</scene>, <scene name='pdbligand=TP2:'>TP2</scene> and <scene name='pdbligand=GOL:'>GOL</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [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] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1F4C OCA].

==Reference==

Line 14:

[[Category: Single protein]]

[[Category: Thymidylate synthase]]

[[Category: Braisted, A.C.]]

[[Category: Braisted, A C.]]

[[Category: Erlanson, D.A.]]

[[Category: Erlanson, D A.]]

[[Category: Gordon, E.]]

[[Category: Randal, M.]]

[[Category: Raphael, D.R.]]

[[Category: Raphael, D R.]]

[[Category: Stroud, R.M.]]

[[Category: Stroud, R M.]]

[[Category: Wells, J.A.]]

[[Category: Wells, J A.]]

[[Category: GOL]]

[[Category: SO4]]

Line 26:

[[Category: e. coli thymidylate synthase modified at cysteine 146]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Tue Nov 20 14~~:36:~~07 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:34:49 2008''

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@@ Line 1: / Line 1: @@
-[[Image:1f4c.jpg|left|200px]]<br /><applet load="1f4c" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1f4c.jpg|left|200px]]<br /><applet load="1f4c" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1f4c, resolution 2.0&Aring;" />
 '''CRYSTAL STRUCTURE OF E. COLI THYMIDYLATE SYNTHASE COVALENTLY MODIFIED AT C146 WITH N-[TOSYL-D-PROLINYL]AMINO-ETHANETHIOL'''<br />
 ==Overview==
-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.
+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 Structure==
-F4C is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with SO4, TP2 and GOL as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [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] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1F4C OCA].
+F4C is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with <scene name='pdbligand=SO4:'>SO4</scene>, <scene name='pdbligand=TP2:'>TP2</scene> and <scene name='pdbligand=GOL:'>GOL</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [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] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1F4C OCA].
 ==Reference==
@@ Line 14: / Line 14: @@
 [[Category: Single protein]]
 [[Category: Thymidylate synthase]]
-[[Category: Braisted, A.C.]]
+[[Category: Braisted, A C.]]
-[[Category: Erlanson, D.A.]]
+[[Category: Erlanson, D A.]]
 [[Category: Gordon, E.]]
 [[Category: Randal, M.]]
-[[Category: Raphael, D.R.]]
+[[Category: Raphael, D R.]]
-[[Category: Stroud, R.M.]]
+[[Category: Stroud, R M.]]
-[[Category: Wells, J.A.]]
+[[Category: Wells, J A.]]
 [[Category: GOL]]
 [[Category: SO4]]
@@ Line 26: / Line 26: @@
 [[Category: e. coli thymidylate synthase modified at cysteine 146]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 14:36:07 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:34:49 2008''