1h42: Difference between revisions

Revision as of 18:24, 2 May 2008

FERREDOXIN:NADP+ REDUCTASE MUTANT WITH THR 155 REPLACED BY GLY, ALA 160 REPLACED BY THR AND LEU 263 REPLACED BY PRO (T155G-A160T-L263P)

OverviewOverview

Previous studies indicated that the determinants of coenzyme specificity in ferredoxin-NADP+ reductase (FNR) from Anabaena are situated in the 2'-phosphate (2'-P) NADP+ binding region, and also suggested that other regions must undergo structural rearrangements of the protein backbone during coenzyme binding. Among the residues involved in such specificity could be those located in regions where interaction with the pyrophosphate group of the coenzyme takes place, namely loops 155-160 and 261-268 in Anabaena FNR. In order to learn more about the coenzyme specificity determinants, and to better define the structural basis of coenzyme binding, mutations in the pyrophosphate and 2'-P binding regions of FNR have been introduced. Modification of the pyrophosphate binding region, involving residues Thr-155, Ala-160, and Leu-263, indicates that this region is involved in determining coenzyme specificity and that selected alterations of these positions produce FNR enzymes that are able to bind NAD+. Thus, our results suggest that slightly different structural rearrangements of the backbone chain in the pyrophosphate binding region might determine FNR specificity for the coenzyme. Combined mutations at the 2'-P binding region, involving residues Ser-223, Arg-224, Arg-233, and Tyr-235, in combination with the residues mentioned above in the pyrophosphate binding region have also been carried out in an attempt to increase the FNR affinity for NAD+/H. However, in most cases the analyzed mutants lost the ability for NADP+/H binding and electron transfer, and no major improvements were observed with regard to the efficiency of the reactions with NAD+/H. Therefore, our results confirm that determinants for coenzyme specificity in FNR are also situated in the pyrophosphate binding region and not only in the 2'-P binding region. Such observations also suggest that other regions of the protein, yet to be identified, might also be involved in this process.

About this StructureAbout this Structure

1H42 is a Single protein structure of sequence from Anabaena sp.. Full crystallographic information is available from OCA.

ReferenceReference

Involvement of the pyrophosphate and the 2'-phosphate binding regions of ferredoxin-NADP+ reductase in coenzyme specificity., Tejero J, Martinez-Julvez M, Mayoral T, Luquita A, Sanz-Aparicio J, Hermoso JA, Hurley JK, Tollin G, Gomez-Moreno C, Medina M, J Biol Chem. 2003 Dec 5;278(49):49203-14. Epub 2003 Sep 18. PMID:14500716 Page seeded by OCA on Fri May 2 18:24:23 2008

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OCA

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 [[Image:1h42.jpg|left|200px]]
- {{Structure
+<!--
-|PDB= 1h42 |SIZE=350|CAPTION= <scene name='initialview01'>1h42</scene>, resolution 2.15&Aring;
+The line below this paragraph, containing "STRUCTURE_1h42", creates the "Structure Box" on the page.
-|SITE= <scene name='pdbsite=AC1:So4+Binding+Site+For+Chain+A'>AC1</scene>
+You may change the PDB parameter (which sets the PDB file loaded into the applet)
-|LIGAND= <scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>
+or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
-|ACTIVITY= <span class='plainlinks'>[http://en.wikipedia.org/wiki/Ferredoxin--NADP(+)_reductase Ferredoxin--NADP(+) reductase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.18.1.2 1.18.1.2] </span>
+or leave the SCENE parameter empty for the default display.
-|GENE=
+-->
-|DOMAIN=
+{{STRUCTURE_1h42|  PDB=1h42  |  SCENE=  }}
-|RELATEDENTRY=
-|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1h42 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1h42 OCA], [http://www.ebi.ac.uk/pdbsum/1h42 PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1h42 RCSB]</span>
-}}
 '''FERREDOXIN:NADP+ REDUCTASE MUTANT WITH THR 155 REPLACED BY GLY, ALA 160 REPLACED BY THR AND LEU 263 REPLACED BY PRO (T155G-A160T-L263P)'''
@@ Line 24: / Line 21: @@
 Involvement of the pyrophosphate and the 2'-phosphate binding regions of ferredoxin-NADP+ reductase in coenzyme specificity., Tejero J, Martinez-Julvez M, Mayoral T, Luquita A, Sanz-Aparicio J, Hermoso JA, Hurley JK, Tollin G, Gomez-Moreno C, Medina M, J Biol Chem. 2003 Dec 5;278(49):49203-14. Epub 2003 Sep 18. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/14500716 14500716]
 [[Category: Anabaena sp.]]
-[[Category: Ferredoxin--NADP(+) reductase]]
 [[Category: Single protein]]
 [[Category: Gomez-Moreno, C.]]
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 [[Category: Medina, M.]]
 [[Category: Sanz-Aparicio, J.]]
-[[Category: fad]]
+[[Category: Fad]]
-[[Category: flavoprotein]]
+[[Category: Flavoprotein]]
-[[Category: fnr]]
+[[Category: Fnr]]
-[[Category: nadp]]
+[[Category: Nadp]]
-[[Category: nadp reductase]]
+[[Category: Nadp reductase]]
-[[Category: oxidoreductase]]
+[[Category: Oxidoreductase]]
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May  2 18:24:23 2008''
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 20:57:14 2008''