User:Yana Fedotova/Sandbox 1: Difference between revisions

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== Background Information ==
== Background Information ==
''
''
PrrA is a DNA binding protein and a essential part of the two-component signal transduction regulatory system, PrrBA.  ''Rhodobacter capsulatus'' contains RegA considered a homologue to PrrA protein in ''R. sphaeroides''.  PrrA is proposed to be a master regulator/response regulator involved in oxygen regulation of photosynthesis genes expression responding to changes to the rate of electron flow as means to maintain redox balance, such as a balance between energy production and energy consumption in the cell <ref> Zeilstra-Ryalls, J. H. and S. Kaplan.  2004.  Oxygen Intervention in the Regulation of Gene Expression: The Photosynthetic Bacterial Paradigm.  Cellular and Molecular Life Sciences 61:417-436.</ref>.  PrrA is thought to play an important function in carbon dioxide and nitrogen fixations, photosynthesis as well as proton oxidation and uptake <ref> Eraso, J. M., J. H. Roh, X. Zeng, S. J. Callister, M. S. Lipton and S. Kaplan.  2008. Role of Global Transcriptional Regulator PrrA in ''Rhodobacter sphaeroides'' 2.4.1: Combined Transcriptome and Proteome Analysis.  Journal of Bacteriology 190: 4831-4848.</ref>.   
PrrA is a DNA binding protein and a essential part of the two-component signal transduction regulatory system, PrrBA.  ''Rhodobacter capsulatus'' contains RegA considered a homologue to PrrA protein in ''R. sphaeroides''.  PrrA is proposed to be a master regulator/response regulator involved in oxygen regulation of photosynthesis genes expression responding to changes to the rate of electron flow as means to maintain redox balance, such as a balance between energy production and energy consumption in the cell <ref> Zeilstra-Ryalls, J. H. and S. Kaplan.  2004.  Oxygen Intervention in the Regulation of Gene Expression: The Photosynthetic Bacterial Paradigm.  Cellular and Molecular Life Sciences 61:417-436.</ref>.  PrrA is thought to play an important function in carbon dioxide and nitrogen fixations, photosynthesis as well as proton oxidation and uptake <ref> Eraso, J. M., J. H. Roh, X. Zeng, S. J. Callister, M. S. Lipton and S. Kaplan.  2008. Role of Global Transcriptional Regulator PrrA in ''Rhodobacter sphaeroides'' 2.4.1: Combined Transcriptome and Proteome Analysis.  Journal of Bacteriology 190: 4831-4848.</ref>.   
PrrB is considered a sensor histidine kinase bound to the cytoplasmic membrane that can phosphorylate PrrA when it senses changes the aforementioned changes in the cell.  Interestingly, “1,058 genes were proposed to be regulated out of 4,284 genes as represented on the GeneChip of ''R. sphaeroides'' 2.4.1” by PrrA <ref> Eraso, J. M. and S. Kaplan.  1994.  ''prrA'', a Putative Response Regulator Involved in Oxygen Regulation of Photosynthesis Gene Expression in ''Rhodobacter sphaeroides''.  Journal of Bacteriology 176:32-43.</ref>.
PrrB is considered a sensor histidine kinase bound to the cytoplasmic membrane that can phosphorylate PrrA when it senses changes the aforementioned changes in the cell.  Interestingly, “1,058 genes were proposed to be regulated out of 4,284 genes as represented on the GeneChip of ''R. sphaeroides'' 2.4.1” by PrrA <ref> Eraso, J. M. and S. Kaplan.  1994.  ''prrA'', a Putative Response Regulator Involved in Oxygen Regulation of Photosynthesis Gene Expression in ''Rhodobacter sphaeroides''.  Journal of Bacteriology 176:32-43.</ref>.
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''
== Structure of the Proposed PrrA Protein ==
== Structure of the Proposed PrrA Protein ==
''<applet load='PrrA.pdb' size='300' color='color' frame='true' align='right' caption='3D Image of proposed PrrA protein'/>
<applet load='PrrA.pdb' size='300' color='black' frame='true' align='right' caption='3D Proposed Image of PrrA Protein'/>
 
<scene name='User:Yana_Fedotova/Sandbox_1/Secondary_structure_-_PrrA/1'>Secondary Structure</scene>
 
<scene name='User:Yana_Fedotova/Sandbox_1/Polar_regions/2'>Polar/Hydrophobic Regions</scene>
 
<scene name='User:Yana_Fedotova/Sandbox_1/N_to_c_rainbow/1'>Amino Terminus to Carboxy Terminus</scene>
 
{{Template:ColorKey_Amino2CarboxyRainbow}}
 
http://ca.expasy.org/tools/protparam.html
http://ca.expasy.org/tools/protparam.html


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- RESIDUE VARIETY: The residues variety at each position of the multiple sequence alignment.
- RESIDUE VARIETY: The residues variety at each position of the multiple sequence alignment.
Phylogenetic Tree of PrrA [http://consurf.tau.ac.il/results/1240259643/treeView.html]


   POS   SEQ     COLOR       RESIDUE VARIETY (Normalized)                        
   POS   SEQ     COLOR       RESIDUE VARIETY (Normalized)                        
Line 297: Line 308:
  121   P     4* E,K,N,P
  121   P     4* E,K,N,P


1umq [http://proteopedia.org/wiki/index.php/1umq]


''
== Phylogenetic Tree of PrrA [http://consurf.tau.ac.il/results/1240259643/treeView.html] ==
''
''
== 1umq [http://proteopedia.org/wiki/index.php/1umq] ==
''


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ANIMATED IMAGE CONDTRUCTION:
ANIMATED IMAGE CONSTRUCTION:


1. Go to the POLYVIEW 3D homepage, http://polyview.cchmc.org/polyview3d.html
1. Go to the POLYVIEW 3D homepage, http://polyview.cchmc.org/polyview3d.html
Line 336: Line 340:
4. You can edit your protein by using the scene authoring tools after loading the applet.
4. You can edit your protein by using the scene authoring tools after loading the applet.
''  
''  
== Page Editor ==  
== Page Creator ==  
''  
''  
Yana Fedotova, email: yana.fedotova@gmail.com, yfedot@bgsu.edu
Yana Fedotova, email: yana.fedotova@gmail.com, yfedot@bgsu.edu
Line 343: Line 347:
''  
''  
Dr. Jill Zeilstra-Ryalls, Susana Retamal and Adam Meade
Dr. Jill Zeilstra-Ryalls, Susana Retamal and Adam Meade
<scene name='1umq/Prra_model/4'>PrrA Model</scene>

Revision as of 17:40, 1 May 2009

Photosynthesis Response Regulator PrrA of Rhodobacter sphaeroidesPhotosynthesis Response Regulator PrrA of Rhodobacter sphaeroides

Background InformationBackground Information

PrrA is a DNA binding protein and a essential part of the two-component signal transduction regulatory system, PrrBA. Rhodobacter capsulatus contains RegA considered a homologue to PrrA protein in R. sphaeroides. PrrA is proposed to be a master regulator/response regulator involved in oxygen regulation of photosynthesis genes expression responding to changes to the rate of electron flow as means to maintain redox balance, such as a balance between energy production and energy consumption in the cell [1]. PrrA is thought to play an important function in carbon dioxide and nitrogen fixations, photosynthesis as well as proton oxidation and uptake [2]. PrrB is considered a sensor histidine kinase bound to the cytoplasmic membrane that can phosphorylate PrrA when it senses changes the aforementioned changes in the cell. Interestingly, “1,058 genes were proposed to be regulated out of 4,284 genes as represented on the GeneChip of R. sphaeroides 2.4.1” by PrrA [3].

Structure of the Proposed PrrA ProteinStructure of the Proposed PrrA Protein

3D Proposed Image of PrrA Protein

Drag the structure with the mouse to rotate

 Amino Terminus                 Carboxy Terminus 

http://ca.expasy.org/tools/protparam.html

Molecular weight: 20483.5 Da

Number of Amino Acids: 184

Theoretical pI: 6.85

Estimated half-life:

The N-terminal of the sequence considered is M (Met).

The estimated half-life is:

  • 30 hours (mammalian reticulocytes, in vitro).
  • >20 hours (yeast, in vivo).
  • >10 hours (Escherichia coli, in vivo).

Total number of negatively charged residues (Asp + Glu): 28

Total number of positively charged residues (Arg + Lys): 28

Atomic composition: Carbon C 894 Hydrogen H 1468 Nitrogen N 268 Oxygen O 268 Sulfur S 7

Total number of atoms: 2905

Extinction coefficients:

Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.

Ext. coefficient 11585

Abs 0.1% (=1 g/l) 0.566, assuming ALL Cys residues appear as half cystines

Ext. coefficient 11460

Abs 0.1% (=1 g/l) 0.559, assuming NO Cys residues appear as half cystines


Instability index: 55.68

Aliphatic index: 95.49

Grand Average of Hydropathicity (GRAVY): -0.311

Alpha-helices: 34.78%

Extended (Beta-sheets): 20.65%

Other (Loops): 44.57%

The Full-Length R. sphaeroides PrrA Amino Acid Sequence of 184 ResiduesThe Full-Length R. sphaeroides PrrA Amino Acid Sequence of 184 Residues

1 maedlvfelg adrslllvdd depflkrlak amekrgfvle taqsvaegka iaqarppaya

61 vvdlrledgn gldvvevlre rrpdcrivvl tgygaiatav aavkigatdy lskpadanev

121 thallakges lppppenpms adrvrwehiq riyemcdrnv setarrlnmh rrtlqrilak

181 rspr


Amino Acid Composition
Amino Acid Number present Percentage of total present
Ala (A) 15 13.0%
Arg (R) 10 10.9%
Asn (N) 6 2.7%
Asp (D) 10 7.1%
Cys (C) 1 1.1%
Gln (Q) 5 2.2%
Glu (E) 11 8.2%
Gly (G) 9 4.9%
His (H) 10 1.6%
Ile (I) 3 3.8%
Leu (L) 21 10.9%
Lys (K) 6 4.3%
Met (M) 6 2.7%
Phe (F) 2 1.6%
Pro (P) 8 6.0%
Ser (S) 7 3.8%
Thr (T) 13 3.8%
Trp (W) 2 0.5%
Tyr (Y) 6 2.2%
Val (V) 12 8.7%
Pyl (O) 0 0.0%
Sec (U) 0 0.0%

Evolution of PrrAEvolution of PrrA

Amino Acid Conservation Scores

The following scores assist in examining evolutionary relatedness of the amino acid sequence of PrrA to that of other amino acid sequences.

- POS: The position of the AA in the SEQRES derived sequence.

- SEQ: The SEQRES derived sequence in one letter code.

- 3LATOM: The ATOM derived sequence in three letter code, including the AA's positions as they appear in the PDB file and the chain identifier.

- SCORE: The normalized conservation scores.

- COLOR: The color scale representing the conservation scores (9 - conserved, 1 - variable).

- CONFIDENCE INTERVAL: When using the bayesian method for calculating rates, a confidence interval is assigned to each of the inferred evolutionary conservation scores.

- CONFIDENCE INTERVAL COLORS: When using the bayesian method for calculating rates. The color scale representing the lower and upper bounds of the confidence interval.

- MSA DATA: The number of aligned sequences having an amino acid (non-gapped) from the overall number of sequences at each position.

- RESIDUE VARIETY: The residues variety at each position of the multiple sequence alignment.

Phylogenetic Tree of PrrA [1]

 POS	  SEQ	    		COLOR		      RESIDUE VARIETY (Normalized)	        	               
  1	   D	    		  7		    	D,N,R             
  2	   R	    		  4		    	K,P,Q,R,Y         
  3	   S	    	 	  7			K,N,S,T           
  4	   L	    		  7			A,I,L,V           
  5	   L	    		  8			A,F,H,L           
  6	   L	    		  5			I,L,V             
  7	   V	    		  8			A,I,L,V           
  8	   D	    		  9			D,E               
  9	   D	    		  9			D                 
 10	   D	    		  7			D,E,N             
 11	   E	    		  1			A,D,E,I,K,N,P,Q,R,S,T,V
 12	   P	    	 	  3			A,D,E,G,K,L,N,P,S 
 13	   F	    	 	  7			F,H,I,L,T,V       
 14	   L	    		  6			A,C,G,L,R,S,V     
 15	   K	    		  1			D,E,G,H,K,N,Q,R,T,W
 16	   R	    		  2			A,F,G,I,L,M,R,S,T,V,W,Y
 17	   L	    		  7			F,I,L,M,T,V       
 18	   A	    		  2		    	A,E,G,I,N,Q,R,S,T 
 19	   K	    		  1			A,F,G,I,K,L,Q,R,T,V,Y
 20	   A	    		  6	    		A,C,G,I,L,M,N,R,S,T
 21	   M	    		  8			A,F,I,L,M         
 22	   E	    		  4			A,E,G,K,Q,R,S,T,V 
 23	   K	    		  1			A,D,E,F,G,K,L,M,Q,R,S,T
 24	   R	    		  3			A,D,E,L,M,N,Q,R,S,W
 25	   G	    		  9			G,K               
 26	   F	    		  4			F,H,L,M,Y         
 27	   V	    		  1			A,D,E,K,L,N,Q,R,S,T,V
 28	   L	    		  7			A,C,L,P,T,V       
 29	   E	    		  2			A,D,E,F,H,I,K,L,R,S,T,V,Y
 30	   T	    		  1			A,C,E,H,I,L,M,Q,S,T,V,W
 31	   A	    		  7			A,F,H,T,V,Y       
 32	   Q	    		  1			A,D,E,F,G,H,N,Q,R,S,T,Y
 33	   S	    		  6			D,G,H,L,N,S,T     
 34	   V	    		  6			A,G,L,S,T,V       
 35	   A	    		  1			A,D,E,H,K,L,N,P,Q,R,S,T,V
 36	   E	    		  1			A,D,E,G,M,N,Q,R,S,T
 37	   G	    		  7			A,F,G,L,V         
 38	   K	    		  2			D,E,H,I,K,L,M,R,V,W,Y
 39	   A	    		  1			A,D,E,H,K,L,Q,R,T 
 40	   I	    		  1			A,D,E,F,G,I,K,L,M,Q,S,V,W
 41	   A	    		  1			A,F,G,I,L,M,V     
 42	   Q	    		  1		   	A,E,H,I,K,L,M,N,P,Q,R,S,T,V
 43	   A	    		  2			A,D,E,I,N,P,R,S,T 
 44	   R	    		  1			A,D,E,G,I,K,L,N,Q,R,V,Y
 45	   P	    		  1			A,D,E,F,H,I,K,M,P,Q,R,T,V
 46	   P	    		  4			A,F,G,N,P,R,V,Y   
 47	   A	    		  7			A,D,G,H,K,N,Q,S   
 48	   Y	    		  5			A,C,H,I,L,P,V,Y   
 49	   A	    		  5			A,C,I,L,V         
 50	   V	    		  4			F,I,L,V           
 51	   V	    		  6			C,F,I,L,M,S,T,V   
 52	   D	    		  9			D                 
 53	   L	    		  6			I,L,M,V,W         
 54	   R	    		  7			G,K,M,N,R,T,V,W   
 55	   L	    		  7			L,M               
 56	   E	    		  7			A,D,E,G,P,S       
 57	   D	    		  4			D,E,G,K,N,R       
 58	   G	    		  5			D,E,G,I,K,L,M,T,V 
 59	   N	    		  7			D,N,S,T           
 60	   G	    		  9			A,G               
 61	   L	    		  6			F,I,L,M,V,W       
 62	   D	    		  5			A,D,E,G,H,K,Q,S   
 63	   V	    		  6			A,F,I,L,T,V       
 64	   V	    		  4			A,C,F,I,L,V       
 65	   E	    		  4			A,D,E,H,K,P,Q,R,Y 
 66	   V	    		  1			A,D,E,I,K,M,N,Q,R,T,V,Y
 67	   L	    		  7			I,L,M,V,W         
 68	   R	    		  7			G,K,L,N,Q,R       
 69	   E	    		  1			A,D,E,G,I,K,Q,R,S,V
 70	   R	    		  1		   	A,D,E,H,I,K,L,M,N,Q,R,S,T,V,W
 71	   R	    		  1			A,D,E,G,H,K,N,Q,R,S,Y
 72	   P	    		  5			A,D,I,K,L,P,Q,T,V,Y
 73	   D	    		  3			A,D,E,G,H,L,M,N,Q,R,T,W
 74	   C	    		  3			A,C,F,I,L,M,T,V   
 75	   R	    		  8			A,K,P,R           
 76	   I	    		  7			I,M,S,V           
 77	   V	    		  7			A,I,L,M,V         
 78	   V	    		  6			F,I,L,M,V         
 79	   L	    		  7			I,L,M,V           
 80	   T	    		  9			S,T               
 81	   G	    		  9			A,G,S             
 82	   Y	    		  7			F,H,K,L,Q,R,S,Y   
 83	   G	    		  6			A,D,G,N,S         
 84	   A	    		  5			A,D,E,N,S,T       
 85	   I	    		  6			E,F,I,L,V         
 86	   A	    		  4			A,D,E,F,I,M,P,S,T 
 87	   T	    		  8			A,D,E,H,L,M,N,T   
 88	   A	    		  9			A,E,K,R           
 89	   V	   		  8			I,L,V             
 90	   A	   		  2			A,D,E,F,I,K,L,N,Q,S
 91	   A	   		  9			A,G,T             
 92	   V	   		  6			A,F,I,L,M,S,T,V,Y 
 93	   K	   		  5			A,D,E,K,N,Q,R     
 94	   I	   		  1		   	A,C,D,G,H,I,K,L,M,Q,R,S,T,V
 95	   G	   		  9			G                 
 96	   A	   		  9			A,S,V             
 97	   T	   		  5			A,D,F,L,Q,T,V,Y   
 98	   D	   		  9			D,E,S             
 99	   Y	   		  7			F,Y               
100	   L	   		  7			I,L,M,V           
101	   S	   		  6			A,C,E,I,P,S,T,V   
102	   K	   		  9			K                 
103	   P	   		  9			P                 
104	   A	   		  7			A,C,F,L,V         
105	   D	   		  7			A,D,E,G,H,N,S     
106	   A	   		  4			A,D,F,I,K,L,N,P,T 
107	   N	   		  6			A,D,E,G,K,N,R,T   
108	   E	   		  5			A,D,E,K,N,Q,R,V   
109	   V	   		  6			I,L,M,T,V         
110	   T	   		  1			A,D,I,L,N,Q,R,T,V,Y
111	   H	   		  7			A,E,H,L,N,Q       
112	   A	   		  8			A,L,R,S,T         
113	   L	   		  5			I,L,V             
114	   L	   		  4			E,H,I,K,L,Q,R     
115	   A	   		  6			A,Q,R,S,T         
116	   K	   		  4			A,I,K,L,N,R,V     
117	   G	   		  6			G,L,S,V           
118	   E	   		  8			E,R               
119	   S	   		  6			A,H,R,S           
120	   L	   		  6			G,L,S             
121	   P	   		  4*			E,K,N,P

1umq [2]


ReferenceReference

  1. Zeilstra-Ryalls, J. H. and S. Kaplan. 2004. Oxygen Intervention in the Regulation of Gene Expression: The Photosynthetic Bacterial Paradigm. Cellular and Molecular Life Sciences 61:417-436.
  2. Eraso, J. M., J. H. Roh, X. Zeng, S. J. Callister, M. S. Lipton and S. Kaplan. 2008. Role of Global Transcriptional Regulator PrrA in Rhodobacter sphaeroides 2.4.1: Combined Transcriptome and Proteome Analysis. Journal of Bacteriology 190: 4831-4848.
  3. Eraso, J. M. and S. Kaplan. 1994. prrA, a Putative Response Regulator Involved in Oxygen Regulation of Photosynthesis Gene Expression in Rhodobacter sphaeroides. Journal of Bacteriology 176:32-43.


ANIMATED IMAGE CONSTRUCTION:

1. Go to the POLYVIEW 3D homepage, http://polyview.cchmc.org/polyview3d.html

2. On the submission form, first select 'animation' in the "type of request" section, select the size of the animation to be generated in pixels(here the size is 600), then upload the PDB format protein structure file in the "source of structural data" section.

3. On the "chain color and rendering section" select 'cartoon' and 'secondary structure'.

4. On "advanced structural annotation" section select 'docking models in Capri format'.

5. Any other forms for the animation may be selected by referring to the "Samples" according to the protein structure to be animated.


JMOL IMAGE CONSTRUCTION:

1. First retrieve your protein sequence from http://www.ncbi.nlm.nih.gov/.

2. Go to 3D-JIGSAW page http://bmm.cancerresearchuk.org/~3djigsaw/ and paste the sequence on the submission page. A .pdb format image of your protein will be sent to you on your email which can be opened by RASMOL.

3. Upload this file on Proteopedia and then load the JMol applet for the protein following instructions on the Help:Editing page http://www.proteopedia.org/wiki/index.php/Help:Editing.

4. You can edit your protein by using the scene authoring tools after loading the applet.

Page CreatorPage Creator

Yana Fedotova, email: yana.fedotova@gmail.com, yfedot@bgsu.edu

AcknowledgmentsAcknowledgments

Dr. Jill Zeilstra-Ryalls, Susana Retamal and Adam Meade