1rj7: Difference between revisions
New page: left|200px<br /> <applet load="1rj7" size="450" color="white" frame="true" align="right" spinBox="true" caption="1rj7, resolution 2.30Å" /> '''Crystal structure o... |
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[[Image:1rj7.gif|left|200px]]<br /> | [[Image:1rj7.gif|left|200px]]<br /><applet load="1rj7" size="350" color="white" frame="true" align="right" spinBox="true" | ||
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caption="1rj7, resolution 2.30Å" /> | caption="1rj7, resolution 2.30Å" /> | ||
'''Crystal structure of EDA-A1'''<br /> | '''Crystal structure of EDA-A1'''<br /> | ||
==Overview== | ==Overview== | ||
The protective antigen (PA) moiety of anthrax toxin transports edema | The protective antigen (PA) moiety of anthrax toxin transports edema factor and lethal factor to the cytosol of mammalian cells by a mechanism that depends on its ability to oligomerize and form pores in the endosomal membrane. Previously, some mutated forms of PA, designated dominant negative (DN), were found to coassemble with wild-type PA and generate defective heptameric pore-precursors (prepores). Prepores containing DN-PA are impaired in pore formation and in translocating edema factor and lethal factor across the endosomal membrane. To create a more comprehensive map of sites within PA where a single amino acid replacement can give a DN phenotype, we used automated systems to generate a Cys-replacement mutation for each of the 568 residues of PA63, the active 63-kDa proteolytic fragment of PA. Thirty-three mutations that reduced PA's ability to mediate toxicity at least 100-fold were identified in all four domains of PA63. A majority (22) were in domain 2, the pore-forming domain. Seven of the domain-2 mutations, located in or adjacent to the 2beta6 strand, the 2beta7 strand, and the 2beta10-2beta11 loop, gave the DN phenotype. This study demonstrates the feasibility of high-throughput scanning mutagenesis of a moderate sized protein. The results show that DN mutations cluster in a single domain and implicate 2beta6 and 2beta7 strands and the 2beta10-2beta11 loop in the conformational rearrangement of the prepore to the pore. They also add to the repertoire of mutations available for structure-function studies and for designing new antitoxic agents for treatment of anthrax. | ||
==Disease== | ==Disease== | ||
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==About this Structure== | ==About this Structure== | ||
1RJ7 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http:// | 1RJ7 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1RJ7 OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Ackerly, H.]] | [[Category: Ackerly, H.]] | ||
[[Category: Compaan, D | [[Category: Compaan, D M.]] | ||
[[Category: Dixit, V | [[Category: Dixit, V M.]] | ||
[[Category: Hymowitz, S | [[Category: Hymowitz, S G.]] | ||
[[Category: Starovasnik, M | [[Category: Starovasnik, M A.]] | ||
[[Category: Vos, A | [[Category: Vos, A M.de.]] | ||
[[Category: Yan, M.]] | [[Category: Yan, M.]] | ||
[[Category: beta-bulge]] | [[Category: beta-bulge]] | ||
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[[Category: tnf]] | [[Category: tnf]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:51:26 2008'' | ||
Revision as of 15:51, 21 February 2008
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Crystal structure of EDA-A1
OverviewOverview
The protective antigen (PA) moiety of anthrax toxin transports edema factor and lethal factor to the cytosol of mammalian cells by a mechanism that depends on its ability to oligomerize and form pores in the endosomal membrane. Previously, some mutated forms of PA, designated dominant negative (DN), were found to coassemble with wild-type PA and generate defective heptameric pore-precursors (prepores). Prepores containing DN-PA are impaired in pore formation and in translocating edema factor and lethal factor across the endosomal membrane. To create a more comprehensive map of sites within PA where a single amino acid replacement can give a DN phenotype, we used automated systems to generate a Cys-replacement mutation for each of the 568 residues of PA63, the active 63-kDa proteolytic fragment of PA. Thirty-three mutations that reduced PA's ability to mediate toxicity at least 100-fold were identified in all four domains of PA63. A majority (22) were in domain 2, the pore-forming domain. Seven of the domain-2 mutations, located in or adjacent to the 2beta6 strand, the 2beta7 strand, and the 2beta10-2beta11 loop, gave the DN phenotype. This study demonstrates the feasibility of high-throughput scanning mutagenesis of a moderate sized protein. The results show that DN mutations cluster in a single domain and implicate 2beta6 and 2beta7 strands and the 2beta10-2beta11 loop in the conformational rearrangement of the prepore to the pore. They also add to the repertoire of mutations available for structure-function studies and for designing new antitoxic agents for treatment of anthrax.
DiseaseDisease
Known diseases associated with this structure: Ectodermal dysplasia-1, anhidrotic OMIM:[300451], Hypodontia, X-linked OMIM:[300451]
About this StructureAbout this Structure
1RJ7 is a Single protein structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.
ReferenceReference
Mapping dominant-negative mutations of anthrax protective antigen by scanning mutagenesis., Mourez M, Yan M, Lacy DB, Dillon L, Bentsen L, Marpoe A, Maurin C, Hotze E, Wigelsworth D, Pimental RA, Ballard JD, Collier RJ, Tweten RK, Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):13803-8. Epub 2003 Nov 17. PMID:14623961
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