1o8y
From Proteopedia
(New page: 200px<br /><applet load="1o8y" size="450" color="white" frame="true" align="right" spinBox="true" caption="1o8y" /> '''SOLUTION STRUCTURE OF SFTI-1(6,5), AN ACYCLI...) |
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'''SOLUTION STRUCTURE OF SFTI-1(6,5), AN ACYCLIC PERMUTANT OF THE PROTEINASE INHIBITOR SFTI-1, TRANS-TRANS-TRANS CONFORMER (TT-A)'''<br /> | '''SOLUTION STRUCTURE OF SFTI-1(6,5), AN ACYCLIC PERMUTANT OF THE PROTEINASE INHIBITOR SFTI-1, TRANS-TRANS-TRANS CONFORMER (TT-A)'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The most potent known naturally occurring Bowman-Birk inhibitor, sunflower | + | The most potent known naturally occurring Bowman-Birk inhibitor, sunflower trypsin inhibitor-1 (SFTI-1), is a bicyclic 14-amino acid peptide from sunflower seeds comprising one disulfide bond and a cyclic backbone. At present, little is known about the cyclization mechanism of SFTI-1. We show here that an acyclic permutant of SFTI-1 open at its scissile bond, SFTI-1[6,5], also functions as an inhibitor of trypsin and that it can be enzymatically backbone-cyclized by incubation with bovine beta-trypsin. The resulting ratio of cyclic SFTI-1 to SFTI-1[6,5] is approximately 9:1 regardless of whether trypsin is incubated with SFTI-1[6,5] or SFTI-1. Enzymatic resynthesis of the scissile bond to form cyclic SFTI-1 is a novel mechanism of cyclization of SFTI-1[6,5]. Such a reaction could potentially occur on a trypsin affinity column as used in the original isolation procedure of SFTI-1. We therefore extracted SFTI-1 from sunflower seeds without a trypsin purification step and confirmed that the backbone of SFTI-1 is indeed naturally cyclic. Structural studies on SFTI-1[6,5] revealed high heterogeneity, and multiple species of SFTI-1[6,5] were identified. The main species closely resembles the structure of cyclic SFTI-1 with the broken binding loop able to rotate between a cis/trans geometry of the I7-P8 bond with the cis conformer being similar to the canonical binding loop conformation. The non-reactive loop adopts a beta-hairpin structure as in cyclic wild-type SFTI-1. Another species exhibits an iso-aspartate residue at position 14 and provides implications for possible in vivo cyclization mechanisms. |
==About this Structure== | ==About this Structure== | ||
| - | 1O8Y is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/ ]. Full crystallographic information is available from [http:// | + | 1O8Y is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/ ]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1O8Y OCA]. |
==Reference== | ==Reference== | ||
Enzymatic cyclization of a potent bowman-birk protease inhibitor, sunflower trypsin inhibitor-1, and solution structure of an acyclic precursor peptide., Marx UC, Korsinczky ML, Schirra HJ, Jones A, Condie B, Otvos L Jr, Craik DJ, J Biol Chem. 2003 Jun 13;278(24):21782-9. Epub 2003 Mar 5. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=12621047 12621047] | Enzymatic cyclization of a potent bowman-birk protease inhibitor, sunflower trypsin inhibitor-1, and solution structure of an acyclic precursor peptide., Marx UC, Korsinczky ML, Schirra HJ, Jones A, Condie B, Otvos L Jr, Craik DJ, J Biol Chem. 2003 Jun 13;278(24):21782-9. Epub 2003 Mar 5. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=12621047 12621047] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Craik, D | + | [[Category: Craik, D J.]] |
| - | [[Category: Marx, U | + | [[Category: Marx, U C.]] |
[[Category: acyclic permutant]] | [[Category: acyclic permutant]] | ||
[[Category: bowman-birk inhibitor]] | [[Category: bowman-birk inhibitor]] | ||
[[Category: sfti-1]] | [[Category: sfti-1]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:14:48 2008'' |
Revision as of 12:14, 21 February 2008
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SOLUTION STRUCTURE OF SFTI-1(6,5), AN ACYCLIC PERMUTANT OF THE PROTEINASE INHIBITOR SFTI-1, TRANS-TRANS-TRANS CONFORMER (TT-A)
Overview
The most potent known naturally occurring Bowman-Birk inhibitor, sunflower trypsin inhibitor-1 (SFTI-1), is a bicyclic 14-amino acid peptide from sunflower seeds comprising one disulfide bond and a cyclic backbone. At present, little is known about the cyclization mechanism of SFTI-1. We show here that an acyclic permutant of SFTI-1 open at its scissile bond, SFTI-1[6,5], also functions as an inhibitor of trypsin and that it can be enzymatically backbone-cyclized by incubation with bovine beta-trypsin. The resulting ratio of cyclic SFTI-1 to SFTI-1[6,5] is approximately 9:1 regardless of whether trypsin is incubated with SFTI-1[6,5] or SFTI-1. Enzymatic resynthesis of the scissile bond to form cyclic SFTI-1 is a novel mechanism of cyclization of SFTI-1[6,5]. Such a reaction could potentially occur on a trypsin affinity column as used in the original isolation procedure of SFTI-1. We therefore extracted SFTI-1 from sunflower seeds without a trypsin purification step and confirmed that the backbone of SFTI-1 is indeed naturally cyclic. Structural studies on SFTI-1[6,5] revealed high heterogeneity, and multiple species of SFTI-1[6,5] were identified. The main species closely resembles the structure of cyclic SFTI-1 with the broken binding loop able to rotate between a cis/trans geometry of the I7-P8 bond with the cis conformer being similar to the canonical binding loop conformation. The non-reactive loop adopts a beta-hairpin structure as in cyclic wild-type SFTI-1. Another species exhibits an iso-aspartate residue at position 14 and provides implications for possible in vivo cyclization mechanisms.
About this Structure
1O8Y is a Single protein structure of sequence from [1]. Full crystallographic information is available from OCA.
Reference
Enzymatic cyclization of a potent bowman-birk protease inhibitor, sunflower trypsin inhibitor-1, and solution structure of an acyclic precursor peptide., Marx UC, Korsinczky ML, Schirra HJ, Jones A, Condie B, Otvos L Jr, Craik DJ, J Biol Chem. 2003 Jun 13;278(24):21782-9. Epub 2003 Mar 5. PMID:12621047
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