2nwd
From Proteopedia
(New page: 200px<br /> <applet load="2nwd" size="450" color="white" frame="true" align="right" spinBox="true" caption="2nwd, resolution 1.04Å" /> '''Structure of chemic...) |
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| - | [[Image:2nwd.gif|left|200px]]<br /> | + | [[Image:2nwd.gif|left|200px]]<br /><applet load="2nwd" size="350" color="white" frame="true" align="right" spinBox="true" |
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caption="2nwd, resolution 1.04Å" /> | caption="2nwd, resolution 1.04Å" /> | ||
'''Structure of chemically synthesized human lysozyme at 1 Angstrom resolution'''<br /> | '''Structure of chemically synthesized human lysozyme at 1 Angstrom resolution'''<br /> | ||
==Overview== | ==Overview== | ||
| - | In this article, we report the total chemical synthesis of human lysozyme. | + | In this article, we report the total chemical synthesis of human lysozyme. Lysozyme serves as a widespread model system in various fields of biochemical research, including protein folding, enzyme catalysis, and amyloidogenesis. The 130-aa wild-type polypeptide chain of the human enzyme was assembled from four polypeptide segments by using native chemical ligation in a fully convergent fashion. Key to the assembly strategy is the application of the recently developed kinetically controlled ligation methodology, which provides efficient control over the ligation of two peptide (alpha)thioesters to yield a unique product. This result enables the facile preparation of a 64-residue peptide (alpha)thioester; this segment is joined by native chemical ligation to a 66-aa Cys peptide, to yield the target 130-aa polypeptide chain. The synthetic polypeptide chain was folded in vitro into a defined tertiary structure with concomitant formation of four disulfides, as shown by 2D TOCSY NMR spectroscopy. The structure of the synthetic human lysozyme was confirmed by high-resolution x-ray diffraction, giving the highest-resolution structure (1.04 A) observed to date for this enzyme. Synthetic lysozyme was obtained in good yield and excellent purity and had full enzymatic activity. This facile and efficient convergent synthesis scheme will enable preparation of unique chemical analogs of the lysozyme molecule and will prove useful in numerous areas of lysozyme research in the future. |
==Disease== | ==Disease== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 2NWD is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/ ]. Active as [http://en.wikipedia.org/wiki/Lysozyme Lysozyme], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.17 3.2.1.17] Full crystallographic information is available from [http:// | + | 2NWD is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/ ]. Active as [http://en.wikipedia.org/wiki/Lysozyme Lysozyme], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.17 3.2.1.17] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2NWD OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Durek, T.]] | [[Category: Durek, T.]] | ||
| - | [[Category: Kent, S | + | [[Category: Kent, S B.H.]] |
| - | [[Category: Torbeev, V | + | [[Category: Torbeev, V Y.]] |
[[Category: chemical protein synthesis]] | [[Category: chemical protein synthesis]] | ||
[[Category: convergent synthesis]] | [[Category: convergent synthesis]] | ||
[[Category: native chemical ligation]] | [[Category: native chemical ligation]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:11:44 2008'' |
Revision as of 16:11, 21 February 2008
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Structure of chemically synthesized human lysozyme at 1 Angstrom resolution
Contents |
Overview
In this article, we report the total chemical synthesis of human lysozyme. Lysozyme serves as a widespread model system in various fields of biochemical research, including protein folding, enzyme catalysis, and amyloidogenesis. The 130-aa wild-type polypeptide chain of the human enzyme was assembled from four polypeptide segments by using native chemical ligation in a fully convergent fashion. Key to the assembly strategy is the application of the recently developed kinetically controlled ligation methodology, which provides efficient control over the ligation of two peptide (alpha)thioesters to yield a unique product. This result enables the facile preparation of a 64-residue peptide (alpha)thioester; this segment is joined by native chemical ligation to a 66-aa Cys peptide, to yield the target 130-aa polypeptide chain. The synthetic polypeptide chain was folded in vitro into a defined tertiary structure with concomitant formation of four disulfides, as shown by 2D TOCSY NMR spectroscopy. The structure of the synthetic human lysozyme was confirmed by high-resolution x-ray diffraction, giving the highest-resolution structure (1.04 A) observed to date for this enzyme. Synthetic lysozyme was obtained in good yield and excellent purity and had full enzymatic activity. This facile and efficient convergent synthesis scheme will enable preparation of unique chemical analogs of the lysozyme molecule and will prove useful in numerous areas of lysozyme research in the future.
Disease
Known diseases associated with this structure: Amyloidosis, renal OMIM:[153450], Microphthalmia, syndromic 1 OMIM:[309800]
About this Structure
2NWD is a Single protein structure of sequence from [1]. Active as Lysozyme, with EC number 3.2.1.17 Full crystallographic information is available from OCA.
Reference
Convergent chemical synthesis and high-resolution x-ray structure of human lysozyme., Durek T, Torbeev VY, Kent SB, Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):4846-51. Epub 2007 Mar 8. PMID:17360367
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