From Proteopedia
(Difference between revisions)
proteopedia linkproteopedia link
|
|
| Line 1: |
Line 1: |
| - | [[Image:2vc4.gif|left|200px]] | + | {{Seed}} |
| | + | [[Image:2vc4.png|left|200px]] |
| | | | |
| | <!-- | | <!-- |
| Line 9: |
Line 10: |
| | {{STRUCTURE_2vc4| PDB=2vc4 | SCENE= }} | | {{STRUCTURE_2vc4| PDB=2vc4 | SCENE= }} |
| | | | |
| - | '''RICIN A-CHAIN (RECOMBINANT) E177D MUTANT'''
| + | ===RICIN A-CHAIN (RECOMBINANT) E177D MUTANT=== |
| | | | |
| | | | |
| - | ==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.
| + | The line below this paragraph, {{ABSTRACT_PUBMED_17916187}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 17916187 is the PubMed ID number. |
| | + | --> |
| | + | {{ABSTRACT_PUBMED_17916187}} |
| | | | |
| | ==About this Structure== | | ==About this Structure== |
| Line 32: |
Line 36: |
| | [[Category: Protein synthesis inhibitor]] | | [[Category: Protein synthesis inhibitor]] |
| | [[Category: Toxin]] | | [[Category: Toxin]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May 4 18:34:26 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 05:05:54 2008'' |
Revision as of 02:06, 28 July 2008
Template:STRUCTURE 2vc4
RICIN A-CHAIN (RECOMBINANT) E177D MUTANT
Template:ABSTRACT PUBMED 17916187
About this Structure
2VC4 is a Single protein structure of sequence from Ricinus communis. 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
Page seeded by OCA on Mon Jul 28 05:05:54 2008
