User:Sujay Joseph/Sandbox1 Choline Oxidase

From Proteopedia

< User:Sujay Joseph(Difference between revisions)
Jump to: navigation, search
Current revision (16:53, 8 December 2009) (edit) (undo)
 
Line 1: Line 1:
-
<applet load='2jbv' size='300' frame='true' align='right' caption='Insert caption here' />== Abstract ==
+
<applet load='2jbv' size='300' frame='true' align='right' caption='Insert caption here' />== Discussion ==
-
Importance of Several Evolutionarily Preserved Amino Acids in Choline Oxidase.
+
Threonine 463, valine 464, and histidine 466 are evolutionarily preserved(the amino acid sequences are the same)from bacteria to mice in an enzyme that changed drastically. We found that the significant amino acids are Histidine 466,Valine 464, Threonine 463 that are 4.09 Å,5.07 Å,7.67 Å respectively from the active site. Each of the three residues should therefore be of some importance to the stability of the flavin group, seeing as they were evolutionarily conserved. It is evident that His466 acts as a base in the active site of choline oxidase (Chen and Murata, 2002). However, the functions of Thr463 and Val464 are unknown. Therefore, further research would be necessary to determine the functions of all of the amino acids lining the active site. Mutational studies will also help a great deal in our understanding of the enzyme to see what effect mutating any combination of these three amino acids would have on the enzymatic activity of choline oxidase. Knowing the functions of the amino acids in the active site will enable us to devise genetically modified choline oxidase enzymes. If any recombinant enzymes are found to be Were any recombinant enzymes found to be functionally better, then they could be introduced to a population of plants to genetically induce potential resistance to drought. On the other hand, recombinant choline oxidase enzymes that were catalytically slower could be studied for their effect on reducing, if not completely suppressing, the growth of pathogenic bacterial colonies.
-
Choline oxidase is the enzyme that catalyzes the reaction between choline and betaine glycine. Betaine glycine is an osmoprotectant and instrumental in helping plants and bacteria survive dry conditions. Studying the choline oxidase may help in the development of controlling populations of beneficial plants or pathogenic bacteria. The data was analyzed using the BLAST and Rasmol programs. The structure of choline oxidase from Arthrobacter globiformis (bacteria) was compared with the structure of the complimentary protein in Mus musculus (mice), carnitine acetyltransferase (Altschul et al., 2005). There are seven amino acids evolutionarily preserved within the vicinity of the flavin group (between amino acids 460 to 483 of each subunit). Of these seven, three are within 9 Å of the flavin group while the other four are farther away. These three are threonine 463, valine 464, and histadine 466. They are colored blue and are connected with the ligand via white monitor lines. The other four conserved residues are colored cyan, but are too far to really interact with the flavin group (colored cpk). Beta sheets are colored yellow to showcase the secondary structure of the entire subunit. Evolutionary preservation of the Thr463, Val464, and His466 may signify an importance in aiding the function of the flavin group as it relates to the activity of the enzyme. Previous studies indicate that His466 is indeed important in the function of choline oxidase
+
== Works Cited ==
 +
Altschul SF, Wootton JC, Gertz EM, Agarwala R, Morgulis A, Schäffer AA, and Yu YK (2005). Protein
 +
database searches using compositionally adjusted substitution matrices. FEBS J. 272, 5101-5109.
 +
Chen TH and Murata N. (2002). Enhancement of tolerance of abiotic stress by metabolic engineering of
 +
betaines and other compatible solutes. Curr. Opin. Plant Biol. 5, 250-257
 +
Joosten V and van Berkel WJH. (2007). Flavoenzymes. Current Opinion in Chemical Biology, 11:195–202
 +
ncbi.org. (2009). Retrieved November 20, 2009, from Protein Databank: www.ncbi.org
 +
Quaye, O., Lountos, G., Fan, F., Orville, A., & Gadda, G. (2008). Role of Glu312 in Binding and Positioning of
 +
the Substrate for the Hydride. Biochemistry, 47, 243-256.
 +
Rodwazowski KL, Khachatourians GG, and Selvaraj G. (1990). Choline oxidase, a catabolic enzyme in
 +
Arthrobacter pascens, facilitates adaptation to osmotic stress in Escherichia coli. J Bacteriol. 173(2), 472-478
 +
Sakamoto A and Murata N. (2001). The Use of Bacterial Choline Oxidase, a Glycinebetaine – synthesizing
 +
Enzyme, to Create Stress-Resistant Transgenic Plants1. Plant Biology Vol. 125, pp. 180–188

Current revision

Insert caption here

Drag the structure with the mouse to rotate
== Discussion ==

Threonine 463, valine 464, and histidine 466 are evolutionarily preserved(the amino acid sequences are the same)from bacteria to mice in an enzyme that changed drastically. We found that the significant amino acids are Histidine 466,Valine 464, Threonine 463 that are 4.09 Å,5.07 Å,7.67 Å respectively from the active site. Each of the three residues should therefore be of some importance to the stability of the flavin group, seeing as they were evolutionarily conserved. It is evident that His466 acts as a base in the active site of choline oxidase (Chen and Murata, 2002). However, the functions of Thr463 and Val464 are unknown. Therefore, further research would be necessary to determine the functions of all of the amino acids lining the active site. Mutational studies will also help a great deal in our understanding of the enzyme to see what effect mutating any combination of these three amino acids would have on the enzymatic activity of choline oxidase. Knowing the functions of the amino acids in the active site will enable us to devise genetically modified choline oxidase enzymes. If any recombinant enzymes are found to be Were any recombinant enzymes found to be functionally better, then they could be introduced to a population of plants to genetically induce potential resistance to drought. On the other hand, recombinant choline oxidase enzymes that were catalytically slower could be studied for their effect on reducing, if not completely suppressing, the growth of pathogenic bacterial colonies.

Works Cited

Altschul SF, Wootton JC, Gertz EM, Agarwala R, Morgulis A, Schäffer AA, and Yu YK (2005). Protein database searches using compositionally adjusted substitution matrices. FEBS J. 272, 5101-5109. Chen TH and Murata N. (2002). Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes. Curr. Opin. Plant Biol. 5, 250-257 Joosten V and van Berkel WJH. (2007). Flavoenzymes. Current Opinion in Chemical Biology, 11:195–202 ncbi.org. (2009). Retrieved November 20, 2009, from Protein Databank: www.ncbi.org Quaye, O., Lountos, G., Fan, F., Orville, A., & Gadda, G. (2008). Role of Glu312 in Binding and Positioning of the Substrate for the Hydride. Biochemistry, 47, 243-256. Rodwazowski KL, Khachatourians GG, and Selvaraj G. (1990). Choline oxidase, a catabolic enzyme in Arthrobacter pascens, facilitates adaptation to osmotic stress in Escherichia coli. J Bacteriol. 173(2), 472-478 Sakamoto A and Murata N. (2001). The Use of Bacterial Choline Oxidase, a Glycinebetaine – synthesizing Enzyme, to Create Stress-Resistant Transgenic Plants1. Plant Biology Vol. 125, pp. 180–188

Proteopedia Page Contributors and Editors (what is this?)

Sujay Joseph

Retrieved from "http://52.214.119.220/wiki/index.php/User:Sujay_Joseph/Sandbox1_Choline_Oxidase"
Personal tools