1pfp
From Proteopedia
(New page: 200px<br /><applet load="1pfp" size="450" color="white" frame="true" align="right" spinBox="true" caption="1pfp, resolution 2.30Å" /> '''CATHELIN-LIKE MOTIF ...) |
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| - | [[Image:1pfp.jpg|left|200px]]<br /><applet load="1pfp" size=" | + | [[Image:1pfp.jpg|left|200px]]<br /><applet load="1pfp" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1pfp, resolution 2.30Å" /> | caption="1pfp, resolution 2.30Å" /> | ||
'''CATHELIN-LIKE MOTIF OF PROTEGRIN-3'''<br /> | '''CATHELIN-LIKE MOTIF OF PROTEGRIN-3'''<br /> | ||
==Overview== | ==Overview== | ||
| - | A protocol for the quantitative incorporation of both selenomethionine and | + | A protocol for the quantitative incorporation of both selenomethionine and selenocysteine into recombinant proteins overexpressed in Escherichia coli is described. This methodology is based on the use of a suitable cysteine auxotrophic strain and a minimal medium supplemented with selenium-labeled methionine and cysteine. The proteins chosen for these studies are the cathelin-like motif of protegrin-3 and a nucleoside-diphosphate kinase. Analysis of the purified proteins by electrospray mass spectrometry and X-ray crystallography revealed that both cysteine and methionine residues were isomorphously replaced by selenocysteine and selenomethionine. Moreover, selenocysteines allowed the formation of unstrained and stable diselenide bridges in place of the canonical disulfide bonds. In addition, we showed that NDP kinase contains a selenocysteine adduct on Cys122. This novel selenium double-labeling method is proposed as a general approach to increase the efficiency of the MAD technique used for phase determination in protein crystallography. |
==About this Structure== | ==About this Structure== | ||
| - | 1PFP is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Sus_scrofa Sus scrofa]. Full crystallographic information is available from [http:// | + | 1PFP is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Sus_scrofa Sus scrofa]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1PFP OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Dumas, C.]] | [[Category: Dumas, C.]] | ||
[[Category: Hoh, F.]] | [[Category: Hoh, F.]] | ||
| - | [[Category: Sanchez, J | + | [[Category: Sanchez, J F.]] |
| - | [[Category: Strub, J | + | [[Category: Strub, J M.]] |
| - | [[Category: Strub, M | + | [[Category: Strub, M P.]] |
[[Category: diselenide]] | [[Category: diselenide]] | ||
[[Category: pg-3]] | [[Category: pg-3]] | ||
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[[Category: semet]] | [[Category: semet]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:28:16 2008'' |
Revision as of 12:28, 21 February 2008
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CATHELIN-LIKE MOTIF OF PROTEGRIN-3
Overview
A protocol for the quantitative incorporation of both selenomethionine and selenocysteine into recombinant proteins overexpressed in Escherichia coli is described. This methodology is based on the use of a suitable cysteine auxotrophic strain and a minimal medium supplemented with selenium-labeled methionine and cysteine. The proteins chosen for these studies are the cathelin-like motif of protegrin-3 and a nucleoside-diphosphate kinase. Analysis of the purified proteins by electrospray mass spectrometry and X-ray crystallography revealed that both cysteine and methionine residues were isomorphously replaced by selenocysteine and selenomethionine. Moreover, selenocysteines allowed the formation of unstrained and stable diselenide bridges in place of the canonical disulfide bonds. In addition, we showed that NDP kinase contains a selenocysteine adduct on Cys122. This novel selenium double-labeling method is proposed as a general approach to increase the efficiency of the MAD technique used for phase determination in protein crystallography.
About this Structure
1PFP is a Single protein structure of sequence from Sus scrofa. Full crystallographic information is available from OCA.
Reference
Selenomethionine and selenocysteine double labeling strategy for crystallographic phasing., Strub MP, Hoh F, Sanchez JF, Strub JM, Bock A, Aumelas A, Dumas C, Structure. 2003 Nov;11(11):1359-67. PMID:14604526
Page seeded by OCA on Thu Feb 21 14:28:16 2008
Categories: Single protein | Sus scrofa | Aumelas, A. | Bock, A. | Dumas, C. | Hoh, F. | Sanchez, J F. | Strub, J M. | Strub, M P. | Diselenide | Pg-3 | Secys | Semet
