2vc3
From Proteopedia
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==About this Structure== | ==About this Structure== | ||
| - | 2VC3 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Ricinus_communis Ricinus communis] with <scene name='pdbligand=ACT:'>ACT</scene> and <scene name='pdbligand=SO4:'>SO4</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/rRNA_N-glycosylase rRNA N-glycosylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.2.22 3.2.2.22] Known structural/functional Sites: <scene name='pdbsite=AC1:Act Binding Site For Chain A'>AC1</scene>, <scene name='pdbsite=AC2:So4 Binding Site For Chain A'>AC2</scene> and <scene name='pdbsite=AC3:So4 Binding Site For Chain A'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VC3 OCA]. | + | 2VC3 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Ricinus_communis Ricinus communis] with <scene name='pdbligand=ACT:'>ACT</scene> and <scene name='pdbligand=SO4:'>SO4</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/rRNA_N-glycosylase rRNA N-glycosylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.2.22 3.2.2.22] Known structural/functional Sites: <scene name='pdbsite=AC1:Act+Binding+Site+For+Chain+A'>AC1</scene>, <scene name='pdbsite=AC2:So4+Binding+Site+For+Chain+A'>AC2</scene> and <scene name='pdbsite=AC3:So4+Binding+Site+For+Chain+A'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VC3 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: toxin]] | [[Category: toxin]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Feb 3 10:51:07 2008'' |
Revision as of 08:51, 3 February 2008
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RICIN A-CHAIN (RECOMBINANT) E177D MUTANT WITH A BOUND ACETATE
Overview
Ricin is a heterodimeric plant protein that is potently toxic to mammalian, cells. Toxicity results from the catalytic depurination of eukaryotic, ribosomes by ricin toxin A chain (RTA) that follows toxin endocytosis to, and translocation across, the endoplasmic reticulum membrane. To, ultimately identify proteins required for these later steps in the entry, process, it will be useful to express the catalytic subunit within the, endoplasmic reticulum of yeast cells in a manner that initially permits, cell growth. A subsequent switch in conditions to provoke innate toxin, action would permit only those strains containing defects in genes, normally essential for toxin retro-translocation, refolding or degradation, to survive. As a route to such a screen, several RTA mutants with reduced, catalytic activity have previously been isolated. Here we report the use, of Saccharomyces cerevisiae to isolate temperature-dependent mutants of, endoplasmic reticulum-targeted RTA. Two such toxin mutants with opposing, phenotypes were isolated. One mutant RTA (RTAF108L/L151P) allowed the, yeast cells that express it to grow at 37 degrees C, whereas the same, cells did not grow at 23 degrees C. Both mutations were required for, temperature-dependent growth. The second toxin mutant (RTAE177D) allowed, cells to grow at 23 degrees C but not at 37 degrees C. Interestingly, RTAE177D has been previously reported to have reduced catalytic activity, but this is the first demonstration of a temperature-sensitive phenotype., To provide a more detailed characterization of these mutants we have, investigated their N-glycosylation, stability, catalytic activity and, where appropriate, a three-dimensional structure. The potential utility of, these mutants is discussed.
About this Structure
2VC3 is a Single protein structure of sequence from Ricinus communis with and as ligands. Active as rRNA N-glycosylase, with EC number 3.2.2.22 Known structural/functional Sites: , and . Full crystallographic information is available from OCA.
Reference
The isolation and characterization of temperature-dependent ricin A chain molecules in Saccharomyces cerevisiae., Allen SC, Moore KA, Marsden CJ, Fulop V, Moffat KG, Lord JM, Ladds G, Roberts LM, FEBS J. 2007 Nov;274(21):5586-99. Epub 2007 Oct 4. PMID:17916187
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