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{{Sandbox_Reserved_Butler_CH462_Sp2015_#}}<!-- PLEASE ADD YOUR CONTENT BELOW HERE -->
{{Sandbox_Reserved_Butler_CH462_Sp2015_#}}<!-- PLEASE ADD YOUR CONTENT BELOW HERE -->
==Your Protein Name here==
<StructureSection load='1stp' size='340' side='right' caption='Caption for this structure' scene=''>
This is a default text for your page ''''''. Click above on '''edit this page''' to modify. Be careful with the &lt; and &gt; signs.
You may include any references to papers as in: the use of JSmol in Proteopedia <ref>DOI 10.1002/ijch.201300024</ref> or to the article describing Jmol <ref>PMID:21638687</ref> to the rescue.


<StructureSection load='1SJ2' size='340' side='right' caption=' ' scene=''>
== Biological Function ==


==Introduction==  
== Structural Overview ==
[http://en.wikipedia.org/wiki/Mycobacterium_tuberculosis ''Mycobacterium Tuberculosis''] [http://en.wikipedia.org/wiki/Catalase-peroxidase Catalase Peroxidase] (''mt''CP) is a <scene name='69/694238/Homodimer/1'>homodimer</scene> with each [http://en.wikipedia.org/wiki/Monomer monomer] consisting of two [http://en.wikipedia.org/wiki/Protein_domain domains]. The overall structure is stabilized by 703 <scene name='69/694239/Water_molecules/1'>water molecules</scene>.


== Mechanism of Action ==


===Catalase Peroxidases===
== Zinc Ligand(s) ==
Catalase-peroxidases are enzymes that degrade hydrogen peroxide. Catalase converts two equivalents of hydrogen peroxide into water and oxygen via a two-step reaction  cycle in which H<sub>2</sub>0<sub>2</sub> alternately oxidizes and reduces the heme iron at the active site. Within peroxidases, oxidation of heme iron involves H<sub>2</sub>0<sub>2</sub> molecules, similar to that in the catalase-catalyzed reaction. Reduction of the heme iron, however, involves hydrogen donors such as NADH, not a second H<sub>2</sub>0<sub>2</sub> molcule <ref name="three">PMID: 12172540</ref>. Catalase-Peroxidases that have been characterized are either homodimers or homotetramers and contain a single heme ''b'' cofactor at the active site. Usually, the primary structure of the subunit can be divided into two halves that have a high level of sequence similarity, most likely due to a gene duplication event.


==Structure==
== Other Ligands ==
[[Image:DotSTRUCTURE.png|300 px|left|thumb|'''Figure 1.''' Overall structure of [http://www.rcsb.org/pdb/explore/explore.do?structureId=1sJ2 1SJ2], showing that the structure is a [http://en.wikipedia.org/wiki/Protein_dimer homodimer].]]


This is a sample scene created with SAT to <scene name="/12/3456/Sample/1">color</scene> by Group, and another to make <scene name="/12/3456/Sample/2">a transparent representation</scene> of the protein. You can make your own scenes on SAT starting from scratch or loading and editing one of these sample scenes.


The two [http://en.wikipedia.org/wiki/Protein_domain domains] of each monomer are primarily [http://en.wikipedia.org/wiki/Alpha_helix alpha helical] and have similar foldings. The similar foldings suggests that the monomer results from a [http://en.wikipedia.org/wiki/Gene_duplication gene duplication] event; however, the C-terminal domain does not contain the [http://en.wikipedia.org/wiki/Heme_B heme ''b''] prosthetic group, while the
</StructureSection>
<scene name='69/694238/N_terminus/5'>N terminal</scene> does. The [http://en.wikipedia.org/wiki/Active_site active site] is therefore located within the N-terminal domain. The two monomers interact through an interlocking hook formed by the N-terminal domains that stabilizes the formation of the dimer <ref name="one">PMID: 1523184</ref>.
== References ==
 
The N-terminal <scene name='69/694238/N_terminus/2'>hook</scene> is formed through hydrophobic interactions between <scene name='69/694238/N_terminus/6'>residues Tyr-28 and Tyr-197 and residues Trp-38 and Trp-204</scene>. This interlocking loop region is also found in similar conformations of other catalase peroxidase structures such as: [http://www.proteopedia.org/wiki/index.php/1itk ''hm''CP] and ''bp''CP.
 
===Active Site===
[[Image:INH.png|300 px|left|thumb|'''Figure 2.''' Active Site with conserved amino acid residues. The green arrow represents the binding site.]]
 
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There are 6 conserved key active site residues that surround the active site.  These <scene name='69/694238/Active_site/2'>active site</scene> residues are <scene name='69/694238/Active_site/3'>Arg 104, Trp 107, His 108, His 270, Asp 381</scene> <ref name="one"/>.
 
In ''hm''CP, which share 55% and 69% identity with ''mt''CP, the heme is buried inside ''Hm''CP-N, and substrate access to the active site is through a narrow channel that prevents access of a large substrate <ref name="three"/>. The location of the <scene name='69/694238/Active_site/5'>binding site</scene> for  [http://en.wikipedia.org/wiki/Isoniazid isoniazid (INH)] is located near the ''δ meso'' heme edge, about 3.8 Å away from the heme iron. This binding site is found within what is considered to be the usual <scene name='69/694238/Substrate_access_channel/1'>substrate access channel</scene> of peroxidases.  The reaction between INH and the enzyme must occur from interaction in a binding site intended for the natural substrate <ref name="two">PMID: 12628252</ref>.  <scene name='69/694238/Active_site/6'>Asp 137</scene> plays a key role in the activation and binding of INH.  Asp 137 creates energetically favorable interactions due to its ability to make hydrogen-bond interactions between its carboxylic acid side chain and the pyridinyl N1 of INH<ref name="one"/>. 
 
 
 
==Clinical Applications==
===Isoniazid Structure===
[[Image:INH_structure._PNG.PNG|300 px|left|thumb|'''Figure 3.''' Chemical Structure of Isoniazid (INH)]]
===Isoniazid Role===
[http://en.wikipedia.org/wiki/Isoniazid INH] is a [http://en.wikipedia.org/wiki/Prodrug prodrug] susceptible to oxidative reactions catalyzed by KatG <ref name="four">PMID: 16566587</ref>. INH action against mycobacteria requires catalase-peroxidase (KatG) function.
==Mechanism==
===Formation of IN-NAD Adduct===
Activation or oxidation of INH by KatG has recently been measured in the terms of production of an IN-NAD adduct molecule that serves as a tight binding inhibitor of InhA. InhA is an enzyme involved in the biosynthesis of [http://en.wikipedia.org/wiki/Mycolic_acid mycolic acids], which are components of the mycobacterial cell wall. Therefore, inhibition of InhA alone is sufficient enough to inhibit mycolic acid biosynthesis and induce cell lysis after exposure of bacteria to INH (4).
[[Image:INH_mechanism2.PNG|400 px|right|thumb|'''Figure 4.''' Mechanism of adduct formation]]
 
 
 
==Mutations==
Resistance to INH is a continuing problem in the development of effective therapeutic regimes designed to eliminate infections from ''M. tuberculosis''. Resistance to INH is due to deletions or mutations in this KatG catalase peroxidase enzyme. There are many possible mutations in this peroxidase that can play a role in the resistance of INH. The most commonly occurring mutation occurs at <scene name='69/694238/Active_site/7'>Ser 315</scene>. A mutation at this amino acid can result in up to a 200 fold increase in the minimum inhibitory concentration for INH.  Ser 315 has been reported to mutate to asparagine, isoleucine, glycine, and most frequently, threonine.  A S315T mutant has the ability to reduce the affinity of the enzyme for INH by increasing steric hindrance and reducing access to the substrate binding site <ref name="one"/>.  Mutation of Ser315 to a Thr in ''mt''CP results in a loss of the activation to the anti-tuberculosis drug (INH) with no loss of either peroxidase or catalase activity (3). Any of the other mutations at this site, except for glycine, would also increase steric hindrance and decrease the accessibility to the binding site.
 
[[Image:Mutation_locations.png|300 px|left|thumb|'''Figure 5.''' Pink residues represent the location of possible mutations.  Green residues represent the active site. Asp 137 is shown in blue.]]
 
Of the active site residues that are involved in enzyme catalyzed activation of INH, only <scene name='69/694238/Active_site/8'>His 108</scene> has been a site for mutations that can increase resistance to INH. His 108 has been reported to mutate to glutamic acid and [http://en.wikipedia.org/wiki/Glutamine glutamine]. These mutations reduce the affinity for INH but the hydrogen bond donor/acceptor groups of glutamine would still allow INH to bind.  However, glutamine wouldn't be able to act as proton shuttle in the way His 108 does in the enzyme-catalyzed activation pathway. 
 
No known mutants have been reported to occur at <scene name='69/694238/Active_site/6'>Asp 137</scene>, although a few mutants nearby could cause local conformational changes and thereby altering the orientation of the Asp 137 side chain, making it less effective in binding and activation of INH <ref name="one"/>. 
 
 
 
 
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==See Also==
[http://proteopedia.org/wiki/index.php/Category:Katg Category: KatG]
 
[http://proteopedia.org/wiki/index.php/Category:Isoniazid Category: Isoniazid]
 
[http://proteopedia.org/wiki/index.php/Catalase-peroxidase Catalase-peroxidase]
 
[http://proteopedia.org/wiki/index.php/1sj2 KatG]
 
==References==
<references/>
<references/>
==Authors==
Student Contributors: Ellie Hughes and Nicole Zimmerman

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA, Elizabeth Hughes, Nicole Zimmerman, Geoffrey C. Hoops, David Emch, Isobel Bowles, Jack Trittipo
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